I like this- it reminds me of how vast the ocean is. It also shows how the character feels like she is helpless and lacking direction, hoping that someone will tell her where to go.

Page 6 is my favorite of all the pages because this page describes why Native Americans along with other indigenous groups, forgo their native language and culture. It just reminds me of how my grandmother use to say she wasn't native American even though she was literally 100% Native Hopi. She hated her Native heritage and this page really conceptualizes this notion on why Native sometimes forgo their own culture (Jennings 7).

This reminds me of the "Dead Internet Theory" that is slowly becoming reality. If academia also becomes rampant with AI generated content and student submissions are also AI generated then who is the institution meant for? Would it reveal the underlying notions that universities carry with them, like the degree simply being a mask for privilege, their role in maintaining power structures, and reinforcing colonial ways of knowing?

This somehow reminds me of the idea of 'mansplaining' in gender, or say anyone with power explains my experience to me, posing as an expert. And because society views men as more powerful, they are listened to. I am pointing this out because an administrator using AI to look academic is accepted, but say a student who uses AI to make an argument or a person from a non-English-speaking nation makes a post in English on social media is immediately ridiculed. This somehow goes to say that the individual may have to already possess some power to be able to pass off as intellectual when using AI.

This reminds me of Dr. Parker's comment/question to Dr. Bialystok when she joined our class about whether AI is, in fact, a grand plan/conspiracy for control. Dr. Bialystok seemed to reject this idea, stating that she didn't think these young programmers were thinking that far ahead, but I wonder if the same can be said for university spaces. Even if AI isn't an overall conspiracy, perhaps the ways in which it is adopted in certain spaces are more plausible instances of control tactics.

This reminds me of the discussion in class where someone referred to a university investing in an AI assistant or a robot while there was heavy freezing and cuts on spending on personnel and other aspects. I am interested to know, even within the current spending, how it is being distributed among various aspects of post-secondary education.

This reminds me of the impact of "publish or perish". We see so many academics heavily rely on AI generated articles that are put through peer review and edited (hopefully by a human but at this point who knows), then sent out for publication. That fear of falling behind in this sphere has made people so desperate that they feel they need to publish an upward of 12 pieces per year.

This reminds me of Karen Hao's argument of needing more AI "bicycles" as opposed to AI "rockets." Interestingly both were released in 2025. Although this article is opposing AI in academic, this section also does acknowledge that there may may use for it in certain cases, tasks, which avoids this "all or nothing" polarization we see a lot in AI discusions.

This reminds me of Biesta's concept of 'learnification', that shift away from true education to learners as passive consumers of knowledge.

This reminds me of the eternal struggle between quantitative researchers and qualitative researchers, and also the struggles WITHIN a particular methodology, like Phenomenology and Grounded theory, where different "camps" form and sort of entrench themselves in one or another position. The academic instinct to stick to a certain paradigm is endlessly fascinating.

Edit: Kinda funny how I wrote this and then in the Lather (2018) chapter, on page 107 she discusses the same issues. "Frequent references are made to how the tensions between qualitative and quantitative help one “think deeply” about underlying assumptions (Spencer Foundation 2009, 77).

I really liked this idea because it reminds me that students are more likely to enjoy reading when they get to choose books that interest them. Not every student is going to enjoy the same type of book, and that's okay. As a future ELA teacher, I want to create a classroom where reading is something students look forward to, whether they're reading a novel, a graphic novel, or another type of text that keeps them engaged, and at the beginning of the school year, will ask them their interest so I have an idea!

This reminds me that level 3 students can understand the main idea, they still gain understanding when there is visuals and teacher support. These students can understand something, but not catch every detail. As a teacher I will make sure to have clear explanations to help them fully understand the lessons and instruction.

I like this because it reminds me that all students should be working towards the same learning goal. Even if different support is needed, it is important. As a teacher, you can differentiate learning without lowering the expectations.

I picked this because it reminds me not to underestimate ELL students. Even if they can't fully understand or speak English yet, they are still capable of thinking critically and learning new concepts.

I like this because it reminds me that every student learns differently. Not every student can learn from one type of text. Using different genres and providing supports can help students better understand what they read.

I highlighted this because it reminds me that literacy isn't just about reading books. Students also need the opportunities to speak, listen and communicate because those are skills that help strengthen their reading and writing.

This part stood out to me because I think most teachers have moments where they aren't sure what to do. It reminds me that no teacher knows everything, and it's okay to keep learning so you can better help your students.

I highlighted this because it reminds me that students are shaped by many different parts in their lives, not just the language they speak. As a teacher, I want to know my students as individuals instead of making assumptions based on one part of their identity.

I like how this section reminds me that getting to know your students is the first step in effective teaching. Understanding students' language levels, backgrounds and interests help the teacher create lessons that are meaningful and accessible for each of the students.

I enjoyed this section because it emphasizes planing lessons that are accessible before students struggle instead of making the changes afterwards. This reminds me that a good lesson plan design benefits every student, not just the ELL's.

This reminds me that children learn science best by doing and exploring.

Paine believes that power should not be passed down through blood. This reminds me of the Sunni branch of Islam, where they believed the successor of Muhammad should be anyone fit to rule, not just a blood relative.

'Noise' is often something we do not want to hear, something that is disrupting our peace. It reminds me of Butler's (2024) idea of 'gender as a phantasm' in Who's Afraid of Gender, where talking about anything 'queer' can create this sense of fear of loss of their life or world view, religion, etc., which has the result of demonizing the word. This fear then leads to not engaging with the 'noise' or even perceiving it as 'noise'.

Author response:

The following is the authors’ response to the original reviews.

We have carefully addressed the insightful comments provided by the reviewers which thoroughly increased our comprehension of the dynamics of centriole amplification. The manuscript has been revised accordingly and put in the context of the two papers we published since our last submission, showing that MCC differentiation is a genuine cell cycle variant. A point by point answer to all reviewer comments is provided below.

Briefly:

We have streamlined terminology and nomenclature in text and figures / better define experimental conditions with nocodazole

We have tested the role of dyneins in the dynamics of centriole amplification

We have done correlative light and electron microscopy on the early stages of centriole amplification

We have analyzed a new single cell RNA seq dataset comparing canonical and MCC cell cycle variants in mouse brain progenitors

Collectively, this allowed us to make a clearer parallel with what occurs during centriole duplication and to demonstrate that centriole biogenesis in the MCC cell cycle is marked by the superimposition of 2 canonical centriole cycles.

We believe the manuscript will interest a broader readership since it now provides more fundamental insights on the mechanism of centriole biogenesis.

Public Reviews:

Reviewer #1 (Public Review):

The manuscript by Boudjema et al. describes the cellular events underlying centriole amplification and apical migration to allow the assembly of hundreds of motile cilia in multi-ciliated cells. For this, they use cell culture models in combination with fixed and live cell imaging using antibody staining and fluorescence from endogenously tagged centriole and deuterostome markers, respectively. The work is largely descriptive and functional analyses are restricted to treatment with the microtubule depolymerizing drug nocodazole. The imaging is state-of-the-art including confocal microscopy, live imaging with optical sectioning and high optical and temporal resolution, as well as super-resolution imaging by ultra-expansion microscopy.

The study does a good job of providing a very detailed description of the dynamics of centrioles and deuterostomes that lead to centriole amplification and apical migration in multiciliated cells. This detailed view was missing in previous work. It also reveals the involvement of microtubules at multiple steps: the formation of a cloud of deuterostome precursors, the nuclear envelope tethering of newly formed centrioles, their separation, and their migration to the apical surface.

It would have been useful to expand the analysis of the role of microtubules by including analyses of the requirement for specific microtubule motors, for a better understanding and additional evidence that microtubule-based transport is involved. A weak point is that there is no visualization of microtubules together with deuterosomes and centrioles at the different steps of centriole amplification and migration, to directly address how these structures may interact with and move along microtubules.

Overall, apart from experimental aspects and since this is largely a descriptive study, the manuscript would benefit from more precise language and a better description of the complex events underlying centriole amplification and movements.

We have streamlined terminology and nomenclature, clarified the description of the complex events, and test the role of dyneins in centriole amplification. Microtubules density in MCC does not allow to extract information from imaging. In addition, we have done correlative light and electron microscopy on the early stages of centriole amplification and analyzed a new single cell RNA seq dataset comparing canonical and MCC cell cycle variants in mouse brain progenitors. We also replied points by points to the reviewer specific comments.

Altogether, our new data allowed to demonstrate that centriole biogenesis in the MCC cell cycle is marked by the superimposition of 2 canonical centriole cycles. We believe the manuscript will interest a broader readership since it now provides more fundamental insights on the mechanism of centriole biogenesis.

Reviewer #2 (Public Review):

This important work will be of interest to centriole and cilia cell biologists. It describes in detail how microtubules control multiple aspects of centriole amplification in brain multiciliated cells. This study provides a greater time-resolved and molecular proteomic mapping of the different steps involved, with or without microtubule disruption. Boudjema et al. show that microtubules are important throughout the centriole amplification process, from the early stages, where the procentrioles emerge from a pericentriolar "nest", through the growth stage where microtubules maintain the perinuclear localisation, to the detachment stage, where microtubules assist in perinuclear disengagement and apical migration. The results are generally well supported by the evidence, but the manuscript would benefit significantly from some heavy editing to introduce more niche terms, standardize abbreviations in text, and labels on figures to help bring the readers, especially non-specialists, along with them - increasing the accessibility of their work.

We thank the reviewer for his/her enthusiasm. We have streamlined terminology and nomenclature and clarified the description of the complex events to increase the accessibility of our work. We also replied points by points to his/her specific comments.

Reviewer #3 (Public Review):

Summary:

In this manuscript, Boudjerna and Balagé et al. aim to elucidate the spatial origin of centriole amplification and the mechanisms behind the formation of an apical-basal body patch in multiciliated cells (MCCs). To this end, they focused on the role of microtubules and developed new tools for spatiotemporal and high-resolution analysis of different stages of centriole amplification, including the centrosome stages, A-stage, G-stage, and MCC-stage. Among these tools, the MEF-MCC cells grown on micropatterns stands out for its versatility as it is not tissue-specific and does not require epithelial cell-to-cell contact for differentiation. Additionally, the CEN2-GFP; mRuby-DEUP1 knock-in mouse model was used to study different stages of centriole amplification in physiological brain MCCs. This model offers an advantage over the previously described CEN2-GFP model by enabling the resolution of early events in centriole amplification through the visualization of DEUP1-positive structures and their dynamics. Finally, the authors leveraged powerful imaging techniques, including super-resolution microscopy, the U-ExM, and high-resolution live cell imaging in order to detect and track centriole amplification, elongation, disengagement, and migration.

By combining the MEF-MCC and knock-in mouse model with spatiotemporal imaging in control and nocodazole-treated cells (treated acutely or chronically), the authors define the sequence of events during centriole amplification, revealing the critical roles of microtubules for the first time. Initially, the centrosome-mediated microtubule network forms, organizing a pericentrosomal nest from which procentrioles and deuterosomes emerge. Their findings indicate the importance of microtubules in recruiting and maintaining pericentriolar material clouds that contain DEUP1, PCNT, SAS6, PLK1, PLK4, and tubulins. Following the amplification stage, the procentrioles mature, leading to cells displaying numerous MTOCs, as demonstrated by regrowth experiments. Mature centrioles then disengage from deuterosomes, attach to the nuclear envelope, and migrate to the apical surface facilitated by microtubules.

Strengths:

The manuscript provides new insights into the regulatory function of microtubules in centriole amplification. Addressing the role of microtubules during different stages of centriole amplification required the development of new tools to study brain MCCs, which will be useful in future studies of MCCs. A notable strength of this manuscript is the authors' thorough and quantitative analysis of highly dynamic processes in MCCs. The precision and detail in describing these dynamic events are impressive. This comprehensive analysis advances our understanding of MCC biology.

Weaknesses:

The role of microtubules and other molecular players during different stages of centriole amplification in brain MCCs can be further studied and strengthened using the tools developed in the manuscript. A more quantitative description of some of the analysis performed in the manuscript is required to strengthen the conclusions.

We thank the reviewer for his/her enthusiasm. We have tested the role of dyneins in the dynamics of centriole amplification, done correlative light and electron microscopy on the early stages of centriole amplification and analyzed a new single cell RNA seq dataset comparing canonical and MCC cell cycle variants in mouse brain progenitors. We also replied points by points to the reviewer specific comments.

Recommendations for the authors:

As you will see, all reviewers felt that the analyses of the involvement of microtubules should be strengthened by including controls and additional experiments. Also, they agree that significant text editing would help to improve the manuscript's accessibility and readability.

Specifically, they would suggest (1) streamline terminology and nomenclature in text and figures; (2) better define experimental conditions with nocodazole (concentrations used, effect on microtubules, effect on canonical centriole duplication); and (3), in the absence of other complementary genetic perturbation experiments, add a limitations paragraph in the discussion about conclusions drawn from nocodazole treatment alone.

Reviewer #1 (Recommendations For The Authors):

Main issues:

(1) The authors use variable terminology to describe the same or similar events/structures. For example, in Figure 1 they refer to "centrosome stage" where they observe a pericentrin "cloud", which they later refer to as a "nest". In all other figures the first stage is not referred to as the "centrosome stage" but as the "cloud stage". Again, they also describe the "cloud" as a "nest" occasionally, but not always. In the cartoon, the nest is termed "centrosome cradle". The variable and inconsistent use of terms is confusing and the authors do not provide any explanation for the use of one vs. another.

The text is now corrected. The centrosome stage corresponds to the stage preceding the beginning of centriole amplification in MCC progenitor. The pericentrosomal cloud of centriole and deuterosome elements forms later on, during the amplification A-stage. The formation of this cloud marks the beginning of A-stage, and persists up to G-stage where it dissolves. When we show that the cloud hosts the first stages of centriole biogenesis, we defined it as a “nest”. We do not use anymore the term craddle.

(2) What prompted the authors to use the term "nest"? It gives the impression that they describe aspecific physical entity/structure (also depicted in this way in Figure 3P, with microtubules outside of this structure), but what is the evidence for this?

The cloud is the spatial entity and the term “nest” is used to define a function of this transient compartment. We decided to keep the term “nest” as we now identified it with correlative light and electron microscopy, in addition to U-ExM, and show that the accumulation of centriole and deuterosome elements is accompanied by the formation of immature procentrioles, deprived of MT walls, as well as immature and empty deuterosomes. The scheme with MT outside the cloud/nest is misleading as we see MT organized by the mother centriole. We have now changed this.

(3) The "nest" may simply be a dynamic accumulation of precursor particles around the centrosome, similar to what has been described for centriolar satellites. Rather than proposing a new entity, I suggest testing whether the "nest" particles may colocalize with PCM1 and thus may be related to centriolar satellites. Based on the data, the nest would simply be the centrosomal MTOC that organizes a radial microtubule array on which particles move around its center. In the absence of other evidence, I am not convinced that a new term is needed.

We totally agree with the reviewer: the centrosome, as MTOC, concentrates centriolar and deuterosome components. This cloud is consistently dissolved when MT are depolymerized or dyneins inhibited. So, the physical entity is a “cloud”. We used the term “nest” to propose one function for this cloud which is to form deuterosomes and centrioles, before they move away for maturation. In fact, deuterosome and centriole formation are hindered when the cloud is dissolved. We have tried to edit the text all over the manuscript to make it clearer.

(4) Role of MTs: are microtubules required or do they just facilitate some of the investigated events?

The reason why the role of MT has not been tested yet during centriole amplification is probably because MT not only constitute the cell cytoskeleton on which molecular motors ride to transport cargos or distribute forces, they are also the core component of the structures we are studying. This is why we have tested a range of nocodazole concentrations and used concentrations where MT are perturbed but not entirely depolymerized, allowing centrioles to be produced (Fig. 4 Supplementary 1A-B). This may lead to an underestimation of the role of MT but we cannot study the role of MT on centriole amplification if centrioles cannot be formed.

Does multi-ciliation in these models eventually occur normally under the concentrations and treatment conditions used here? This should be tested and discussed in the context of whether microtubules are indeed required and at what step of the entire process (amplification, migration, ciliogenesis) they may be critical.

We did both chronic and acute treatments.

Chronic treatments were done to test the overall efficiency of centriole amplification when MT (or dyneins) are perturbed. Chronic treatments were used to assess the role of MT (or dyneins) on the global efficiency of centriole and deuterosome formation (number of cells able to amplify, number/size/loading of deuterosomes, final number of centrioles (Fig. 4H-I, Fig. 4 Supplementary 2 B-D). In these chronic treatment, we focused on centriole amplification and not ciliation since it was the scope of this study. Also, we did not take ciliation as a readout of amplification because ciliation is relying on MT polymerization.

Then, we also did acute treatments to test the role of MT (or dyneins) at each stage of amplification (A-amplification, G-growth, D-disengagement, M-migration; Fig. 4, 5, 7, 8 and associated supplementary figures). Since one stage is dependent on the precedent one, this enabled us to decipher the direct role of MT (or dyneins) on each single stage. We have now edited text, methods, legends and pictograms to be clear on whether acute or chronic treatment was done.

(5) Can the authors include control (non-amplifying) progenitors in their analyses? It would be useful to know what the signal and distribution of each specific marker are before differentiation begins (before the cloud stage).

Non amplifying progenitors are analyzed and constitute the so-called “centrosome stage”. We have now precised it and called it the “progenitor stage”.

(6) Figure 2: Again, the terminology is confusing, since the authors describe that DEUP1 forms a "cloud" with centrin during the A stage.

Corrections have been done as explained in point 1.

(7) Description Figure 3: the authors introduce yet another term: "halo" A-stage. Is this the early A stage? Again, this is not explained and confusing. More systematic and consistent description is needed.

Corrections have been done as explained in point 1. The term halos is used un the lab as it was the first term we used in our Nature paper in 2014 in reference to the halo described by Erich Nigg when they overexpressed Plk4. It was an error to use it in the manuscript.

(8) Nocodazole treatments: the used concentrations are quite high.

MCC develop a very dense and stable MT network that is not comparable to cycling cells. MT are very difficult to depolymerize entirely (Fig. 4 Supplementary 1A-B).

(a) To avoid non-specific effects the authors should test what the minimal concentration is that completely depolymerizes microtubules in their cell model and perform analyses at this concentration.

We have of course tested a range of nocodazole concentrations at the beginning of the study (Fig. 4 supplementary 1A-B), and used concentrations where MT are perturbed but not entirely depolymerized, allowing centrioles to be produced (see answer to point 4). In case it was not clear, we refer to this now several time and more clearly in the text and methods.

(b) They should demonstrate depolymerization of microtubules by microtubule staining in the acute and chronic noc treatments and at the different noc concentrations used.

This is, and was, in supplementary material (same, Fig. 4 supplementary 1A).

(c) The authors should demonstrate that the used nocodazole concentrations do not impair normal centriole biogenesis during the cell cycle in these cells; if so, impaired assembly of centriole wall MTs may contribute to the observed effects in Figure 4.

As mentioned in point 8b, we have of course tested a range of nocodazole concentrations at the beginning of the study (Fig. 4 supplementary 1A), and used concentrations where MT are perturbed but not entirely depolymerized, allowing centrioles to be produced (see answer to point 4). The ability of the cells to form centrioles during chronic treatments were always assessed using immunostainings of SAS6 and/or CEN2-GFP signals (now exemplified in Fig. 4 Supplementary 1B). We also did EM analysis on cells treated with the highest doses of nocodazole (Nocodazole 10 uM for 24h) and this showed that centrioles can form with, what seems to be MT walls, in cells totally deprived of cytoplasmic MT fibers (Fig. 4 Supplementary 3-4). However, this does not show that all the cells can, because the number of cells that can be analyzed by EM are not sufficient to conclude. Also, one cannot assess whether MT walls are properly polymerized. However, the absence of MT walls should not change the results of the Figure 4, which are based on DEUP1, SAS6 or CEN2-GFP signals for deuterosomes and centrioles. Also MT depolymerization affects the formation of deuterosomes, which should not be altered by MT wall defects as it is not affected, even when centriole formation is blocked (LoMastro et al., 2024). Last but not least, we now show that blocking dyneins, as a comparable and even greater effect, on the formation of the cloud, deuterosomes and centrioles (Fig. 4C-I and Supplementary Fig. 4), which confirms that MTOC function, rather that MT wall formation, explain the centriole biogenesis alteration shown in Figure 4.

(9) The authors repeatedly refer to the centriole-to-centrosome conversion of amplified centrioles and how this resembles centriole-to-centrosome conversion during the cell cycle. However, they incorrectly claim that this occurs at the G2/M transition. PLK1-dependent modification occurs at this stage, but conversion and PCM recruitment only occur after mitosis (see original work by the Tsou lab, which needs to be cited here).

We agree with the reviewer. We have now added additional data to show clearly that centriole biogenesis, which requires two cell cycles to proceed in cycling cells, is accelerated during the MCC cell cycle variant where the elongation and maturation cycles are superimposed. This is now clearly shown in Fig. 3, 5, 9 and discussed.

(10) Figure 6H-J: the authors claim that at low noc concentration, more D-stage cells showed incomplete disengagement than in controls, but the effect is shown only for the highest 10 µM concentration. Do any eof the phenotypes in Figure 6 also occur at the lowest noc concentration (assuming it depolymerizes MTs)? Again, it is crucial to demonstrate this, to exclude unspecific effects not linked to MT depolymerization.

An error was made on the figure (but not in the legend). In Figure 6, chronic treatments are at 1 or 5 µM. Only acute treatments were done using 10 µM. In both cases, MT are not entirely depolymerized in these experiments (Fig. 4 supplementary 1A).

(11) Disengagement, Figure 7: The authors describe that DEUP1 signal spreads all over the cytoplasm and becomes diffuse during this process, but one cannot see a diffusive signal throughout cells in the figures.

We pushed the contrast to make it clearer but the deuterosomes are still bright at this stage and it is difficult to have both signal clear (now in Fig. 6B). We have also changed the example in video (now video 19) to show it more clearly with DEUP1 channel alone.

(12) Figure 7: localization of disengaged centrioles at microtubule "nodes" is not clear from the images. There are many centrioles and random colocalization may be expected simply based on the high number. Higher resolution and/or magnification and quantification would be needed.

We have edited and now say that centrioles “colocalize” with MT which, since centrioles nucleate MT, seems normal. We agree that it could be random, but given the density of MT, and the number of centrioles, it does not seem opportune to us to quantify. We can just say that we never see centrioles is regions that are deprived of MT.

(13) The term "diffusive" to describe slow centriole movements in Figure 8 suggests that it is not motor or force-dependent, but there is no evidence for that. Movement based on opposing forces could produce a similar result, but would not be considered diffusive.

We agree. We have changed “diffusive” by “diffusive-like”.

(14) The manuscript would greatly benefit from the analysis of some candidate motor activities that may drive the movement and migrations of centrioles in this system. This would support the importance of the microtubule network for the specific steps in these processes, and better define its role beyond "being required". Dynein may be a candidate or minus end-directed kinesins. Since chemical inhibitors are available, these types of experiments would be straightforward.

We formerly tested ciliobrevin but had hard time because of the small stability of the drug. Since our submission to eLife, we tested dynapyrazol and dynarestin and found dynapyrazol very efficient in dissolving the Golgi, a good readout of dynein inhibition. We sought to test the role of dyneins, using dynapyrazol, on (i) the formation of the pericentrosomal cloud in A-stage, (ii) the oscillation of DEUP1+ structures during A-stage, (iii) the number, size, loading of deuterosome, (iv) the final number of centrioles, (v) the migration to the nuclear membrane and (vi) the final apical migration of centrioles. The results are now inserted in main and associated Fig. 4, 5, 7, 8, 9.

(15) Discussion:

"the role microtubules" lacks "of"

This is now edited.

"This lack is..." Lack of what?

This is now edited.

"reflexive link" - meaning of "reflexive" is not clear in this context

We have removed it.

In my opinion, the study does not identify a nest composed of DEUP1, PCNT, and Centrin2; it only shows that these components accumulate as particles around the centrosome, which functions as MTOC. Consequently, it seems that the "nest" does not exist when MT is depolymerized. One could consider the center of the centrosomal MT array as a nest in this context, but there is no evidence of a specific new structure as suggested by the way the term is used in the manuscript.

This is what we want to say: the center of the MT array become a nest in this context. We do not state that there is a specific new structure. We just say that MT and dynein dependent concentration of centriole and deuterosome components exists and that this region nests the birth of centrioles and deuterosomes. Also, this compartment is restricted in time and space, which justifies to use a specific term. The MTOC exists in the progenitor cell, while this compartment, marked by DEUP1, Centrin, PCNT accumulation, appears at the beginning of amplification and grows during A-stage to be dissolved at G-stage when all the deuterosomes and centrioles have moved away.

What is the evidence that "DEUP1 is a centrosomal protein before building deuterosome structures"? It would be good to refer to the specific experiment. Does DEUP1 localize at centrioles also in the absence of microtubules? If not, I would not consider it a centrosomal protein.

We have removed this statement to avoid misinterpretation.

"This reminds the centriole-to-centrosome conversion..." the sentence is missing an "of"; also, again the authors confuse the order of events during the cell cycle, where centrosome conversion occurs after completion of mitosis, not at G2/M transition.

We have removed this statement to avoid misinterpretation. Also, see Point 9.

"microtubule dependent nuclear migration" should be rephrased; it sounds as if the nucleus migrates.

This has been changed

The following discussion of disengagement being linked to association with the nuclear envelope and resembling the process in cycling cells is misleading. In cycling cells movement of centrioles along the nuclear envelope occurs at G2/M and drives centrosome separation (separation of centriole pairs) in preparation for mitosis, not centriole disengagement.

We are now clearer. We compare centriole-loaded deuterosome organization around the nuclear membrane to the migration of new centrosomes during early prophase (Fig. 5F-H, Fig. 5 Supplementary 2G-K).

Regarding the possibility that forces by microtubules generated by the daughter centriole drive disengagement also in cycling cells, I would argue that this is unlikely since the daughter centriole can only nucleate microtubules after disengagement has occurred (and conversion to centrosome/PCM recruitment). Once this happens, it may physically separate the disengaged centrioles, which is a different type of activity. Indeed, originally the term "disengagement" was coined to specifically describe the loss of the perpendicular engagement of daughter centrioles with their mothers (Tsou and Stearns, Nature, 2006).

We have removed this statement to avoid misinterpretation. The perpendicular engagement is difficult to assess on deuterosomes but we do see by live imaging, that attachment changes during D-stage, before centrioles detach clearly from deuterosomes.

"high resolutive" should be "high resolution"

Edit done.

"splitted" should be "split"

Edit done.

"Consistently, when the mitotic oscillator is dis-inhibited and cells enter pseudo-mitotic events, centrioles show clear and rapid cell-cycle like clustering" This sentence is not understandable without further explanation; what does mitotic oscillator refer to? What are pseudo-mitotic events? What is cell cycle-like clustering?

We have removed this statement.

Minor:

(1) Abstract: "Centriole number must be restricted to two..." Since cells are born with two centrioles and have 4 centrioles (2 pairs) when they enter mitosis, this sentence is inaccurate.

The sentence has changed.

(2) Abstract: "reflexive link"; I am not sure what the term "reflexive" refers to?

We have removed this statement to avoid misinterpretation.

(3) Figure 1C, D: it should be described better that the larger magnification panels represent overlays of many cells and what marker they show. This is not obvious since the smaller single-cell panels always show two different markers. Also, it would be more useful to show also single cells in the magnified view. The overlay does not allow us to see if a marker forms a cloud or a single dot, which is as important as the cell-to-cell variation in distribution.

We have clarified this in the text and the legend. The cell-to-cell variation cannot be estimated with the overlay, but the projection from several cells (number precised) allows to see that the signal is confined in a restricted region. Or not. Which is what we wanted to analyze.

Related to the above, the authors say that pericentrin forms a cloud at the top left in panel D, but there is only one confined centrosomal dot in the single-cell panel.

The sentence has changed.

(4) Results, Figure 2F; video 4: The authors claim connection and disconnection of DEUP1 aggregates with centrosomal centrioles; can the authors comment on the spatial resolution including in z in this movie to support this claim? Can they exclude that the structures are in proximity of each other rather than "connected"?

This is a single z-section of 500nm. The resolution in xy is 128nm/pixel. Given the sizes of deuterosomes and a mature centriole, and given the fact that we observed this dynamics in several cells in live, we can state that the structures are connected. This is consistent with deuterosomes frequently observed “kissing” the daughter centriole by EM in the present manuscript (Fig. 2D, Fig. 2 supplementary 3 and 4 and Fig. 4 Supplementary 3-4). One has to look carefully at the daughter centriole (marked “dc”) and span in on the serial sections to see the connected deuterosome (marked by a star): this is at very early stage and therefore it is small. We have not zoomed in since previous manuscript have already described this at later stages with bigger deuterosomes. You can refer to main or supplementary figures in previous manuscripts (Al Jord 2014, Khoury Damaa 2024) where serial sections span the entire deuterosomes and daughter centrioles and show, with nanometric resolution, that both structures are frequently sticked to each others on tens of nanometers.

(5) The term "dynamics" as used in the manuscript should be plural.

It has been used plural, except when for “dynamic microtubules” and “dynamic attachment to the nucleus”, which we think is ok? We have not found any other singular uses in our manuscript.

(6) Figure 5: what does "YL1/2 procentriole intensity" refer to in panel F? This should be the intensity of microtubule asters.

This has been modified.

(7) Figure 6 - supplement 1B: contrary to the claim in the text, one cannot see tight colocalization with the nuclear pore marker. This seems to be a very small subset of particles and even in those cases colocalization is not tight. Also, what is the relevance of nuclear pore colocalization?

We edit and change the phrasing as ‘colocalization with NPC’ is not the good term. What we want to say is that there is a tight connection with the nuclear envelope as shown by the localization of NPC on the same z-section as centrioles. This is why we present a single z, to show that centrioles and NPC are on the same z-plane of 500nm. NPC are stained to outline the nuclear membrane. This is also clearly visible for G-stage centrioles in the XY plane. We have now added an entire z-stack on video 18.

Reviewer #2 (Recommendations For The Authors):

To improve accessibility of their manuscript, we would suggest making the following edits:

(1) Define 'specialist' or 'niche' terms each time you introduce them, such as 'pericentrosomal nest', or 'flower-like structures'.

This has been clarified.

(2) Have a think about abbreviations, again ones that work for people outside the project- this paper uses 'PC' for 'procentriole' but for many 'PC' is 'Parental centriole' or Figure 6J talks about 'D total' or 'D partial', leaves readers confused.

This has been clarified.

(3) Standardize your abbreviations throughout particularly for your treatments- sometimes Noco sometimes, NOCO, or your imaging experiments sometimes Cen-GFP, sometime CEN2-GFP (Figure 7A, D vs. Figure 6) or DEUP1- mRuby, DEUP1-mRuby3 or mRuby3-DEUP1?

We now use Nocodazole or Noco in the text and the figure respectively, CEN2-GFP and mRubyDEUP1.

(4) About 10% of the population, including several key figures in this field, are red-green color blind. Although 4 colour fluorescence is difficult to get right for everyone, choosing palettes (especially for two colour panels) is inclusive. More so, greyscale or inverted monochrome images make it easier for everyone to visualize changes in localization, size, and intensity. Red on black small foci is particularly difficult to discern. For example, Figure 3 - more individual channels in grayscale with arrows to mc, dc, and cilia would be helpful - difficult to distinguish stainings.

We thank the reviewer for this comment and for this recommendation of being more inclusive. We have done the changes.

To improve the conclusions drawn, we suggest some revisions below:

(1) Since the paper really hangs on it, a clearer description of the rationale for when, how long and how much nocodazole treatment was done is needed. The logic currently is difficult to follow seemingly random jumps 10x concentration are used. Microtubules control many aspects of cell biology and could be impacted. For example, I particularly found Figures 6D and H difficult to follow i.e. the timing for 6H seems off.

MCC develop a very dense and stable MT network that is not comparable to cycling cells. MT are very difficult to depolymerize entirely. We have of course tested a range of nocodazole concentrations at the beginning of the study and shown the extent of MT depolymerization under each treatment. We used concentrations where MT are perturbed but not entirely depolymerized, allowing centrioles to be produced (see answer to point 4 reviewer 1). The level of perturbation of MT and consequences on centriole formation at the different timings and doses were done for each experiment and are exemplified in Fig. 4 supplementary 1A-B. This figure was already present in the first version of the manuscript but we have now edited text, methods and pictograms to clarify this.

(2) Perhaps an extension of this point- in general how interdependent are the processes? If there is a defect at the nest stage, how much are the later defects secondary to this, or do MTs genuinely play direct roles at all stages or are these knock-on effects? How do the authors rule this out? Defects in the nest, lead to smaller and more DEUP1+ foci, with defects in concentrating procentriole factors and centrin, which lead to... For example, Figure 4B looks like centrin is reduced upon noco treatment? Does noco treatment affect Cetn2GFP levels globally? Individual channels grayscale would help visualise this better.

See also our answer to reviewer 1 point 8c.

The stages are indeed interdependent. This is why we did both chronic and acute treatments. Chronic treatments were done to test the overall efficiency of centriole amplification when MT are perturbed. We typically used low dose of 1µM because nocodazole remains 48h in the culture medium. Acute treatments were done to test the role of MT at each stage of amplification (A-amplification, G-growth, D-disengagement, M-migration). Most of the acute treatments were done live and nocodazole was applied after the first time point of live monitoring. We used 10µM to have a rapid effect, and because nocodazole remains only several hours in the culture medium. This allowed to monitor the stage “n”, in cells where the stage “n-1” was completed without any drug which allowed to analyze a stage without having perturbed the precedent one.

We now also test the consequences of dynein inhibition using both acute and chronic dynapyrazole treatments. We show that except for centriole migration, dynein inhibition phenocopies MT depolymerization (centriole number, perinuclear organization and disengagement as well as deuterosome number/loading/size).

Nocodazole chronic treatments do affect intensity of CEN2-GFP at G-stage centrioles suggesting an altered A-to-G transition. In D-stage, CEN2-GFP signal seems normal. We now mention this in the text and in the Fig. 4 Supplementary 1B.

(3) The authors nicely show the importance of MTs in the structure of the nest from which procentrioles and DEUP1 positive structures emerge. They suggest this nest may be what supports procentriole generation in the absence of DEUP1 and parental centrioles. Firstly how does this nest look in the absence of DEUP1 and/or parental centrioles (centrinone treatment)? This may be what they are trying to show in Figure 5 Supplement 1 but it currently is very difficult to digest what it is showing relative to controls and whether this is significant in the way it is plotted.

The nest is conserved in the DEUP1KO with or without centrosomal centrioles, as shown by accumulation of Centrin and PCNT at the center of the self-organised MT network (Mercey et al., 2019). This is in fact what motivated our study on the role of MT in centriole amplification. We have edited the legend to precise the quantification done, which is not related to this question. In this quantification, we show that the increased propensity to accumulate PCNT by centriole-loaded deuterosomes between A and G-stage is maintained in the absence of deuterosomes, indicating that centrioles themselves accumulate/recruit PCNT.

(4) Can you do CLEM on DEUP1-Ruby and these early foci at the cloud stage to see if they are visible at the ultrastructural level, relative to procentrioles, microtubules, and other electron-dense structures?

We thank the reviewer for this question. We have done CLEM on the pericentrosomal cloud during very early steps of centriole amplification. This showed that DEUP1 early accumulation at the centrosome corresponds to a region rich in fibro granular aggregates, suggesting that DEUP1 may be translated here, through locally concentrated centriolar sattelites, known to be involved in local translation. Then, small deuterosomes and immature centrioles are formed, within this cloud of sattelites, confirming that the pericentrosomal cloud is a nest for centriole biogenesis (Fig. 2C-D + Fig. 2 Supplementary 2-6 for control and Fig. 4 Supplementary 3-4 for nocodazole treated cells). This also shows that immature deuterosomes are not necessarily round shaped, and can be deprived of centriole loading.

(5) Check the scale bars- see Fig 4E. Check throughout.

Done.

(6) Figure 3 Supplement 1 and 2 don't match the legend and are likely reversed - which one is right?

Done.

(7) Technical issue - I couldn't play videos 6 or 16? Check these work.

Done.

(8) Nomenclature mammalian proteins- mouse or human- should be all caps DEUP1, PLK4, SAS6,etc. Watch your units- space between number and unit.

This has been done.

(9) Many of the graphs involve three biological replicates but why not plot the mean of each of the three experiments and do stats? The number of events measured may conflate the significance. Try using Superplots.

Here is how we proceed: we count the number of occurrence of the phenotype we monitor, and the total number of cells. We apply a X² to test whether there is a significative difference between our replicates in each condition. If not, we pool the number of occurrence of the phenotype we monitor and the total number of cells for the 3 replicates, and for each condition. Finally we apply a X² between the different conditions. This is how we usually proceed to avoid comparing a mean of percentages. This is now explained in the methods.

Minor points:

(1) "DEUP1 is a centrosomal protein and assembles deuterosomes in the pericentrosomal region in brain MCC". I am not sure you have evidence that DEUP1 is a centrosomal protein. You don't seem to study the relationship between centrosomes and DEUP1? Rewrite this title and tone down this claim.

This has been modified.

(2) Why the crossbow micropattern (versus some other shape) - seems very specific but not discussed?

We wanted a shape where centrosome is not localized at the center of mass of the nucleus. Among the corresponding patterns, the crossbow was the one where differentiating cells had less propensity to detach.

(3) Figure 2 - are the foci of DEUP1 at the cloud stage smaller than at A stage? How do they grow? Measure the diameter at cloud stage, just after they leave the cloud and then once they move away from centrosomal cloud and each other. If so, and they do indeed grow in size from the cloud stage to the growth stage which I think your images suggest - do you envision this happening with the gradual addition of DEUP1 rather than fusion?

Early deuterosomes are not easy to detect by light microscopy, because of accumulation of DEUP1 in the cloud. We did CLEM on the cloud of early A-stage cells to resolve the earliest deuterosomes which are often very small (see Fig. 2D, Fig. 2 Supplementary 2-6) suggesting that they grow, either by fusion, which we never observe in our movies at later A-stage, or by accretion of DEUP1. However, by light microscopy, we can detect very early but big deuterosomes, which we see splitting later on into smaller ones. So, we cannot conclude on the mechanism that regulate deuterosome size. This is now discussed in the discussion of the manuscript.

You say in the discussion:

"Consistently, we never observed fusion events of DEUP1 condensates in our time-lapse experiments. More importantly, we did FRAP experiments on endogenously tagged mRuby-DEUP1 in cells at the different stages of centriole amplification, and did not find significant recovery, supporting that centrosomal DEUP1+ foci and deuterosomes are not liquid-like structures (Figure 8 Supplementary 2)." How do you prove there is no fusion of deuterosomes?

It is always difficult to prove the absence of something, we agree! But we did tens of movies with high temporal resolution and never observed fusion events. But, as we say in the previous question, the very early deuterosomes can be very small and we do not distinguish them from the DEUP1+ cloud by live imaging. So at this stage, we cannot say. But later on, during A- or G-stage and when deuterosomes are outside the cloud to be easily observed, we very often observe deuterosomes bumping into each others and stay in close contact for minutes, but then moving away. This, for us, supports the lack of fusion properties. But the question remains open. We now explain this in the manuscript and have added an example in video 28.

If they are getting bigger as I think your imaging suggests from cloud to growth stage, then how is this happening?

MT depolymerisation and dynein inhibition leads to the formation of very small deuterosomes. Dynein inhibition can even lead to a block in the formation of new deuterosomes suggesting that DEUP1 concentration is a crucial parameter for condensation into deuterosomes. Deuterosome growth may happen through oligomerization of DEUP1 molecules allowed by their dyne-independent concentration. Sorokin in 1968 proposed that a supersaturation of deuterosome components may lead to their solid crystallization into deuterosomes. Deuterosome size can also be regulated by a more complex molecular cascade, involving post-translational modifications of DEUP1 or PCM, such as phosphorylations driven by the cell cycle machinery. This would be consistent with the fact that deuterosomes are very big in the absence of CCNO, a cyclin required for entering the MCC cell cycle variant. This will need further investigations.

I'm not sure FRAP actually proves fusion doesn't happen.

Agreed, this is not what we wanted to say, we clarified. The FRAP experiment just suggests that it is not liquid-like.

It is technically difficult to laser ablate individual or only subsets of deuterosomes...

This is what was done but anyway, FRAP does not firmly show that deuterosome compartments are not liquid-like as we now precise.

(4) How do you fix your cells for expansion as you have no preservation of cytoplasmic microtubules? You are saying that there is a "nest" of MTs but beta tubulin ONLY stains the cilia and centriole - why is this? Tyrosinated tubulin on regular confocal shows strong cytoplasmic staining. See Figure 3.

Cytoplasmic microtubules do not preserve well through the expansion process. We did try a few different fixations and pre-extraction methods but they come at a trade-off to preserving centrioles. i.e. we could either preserve cytoplasmic tubes or centrioles but not both with the same processing method.

(5) "PCNT puncta partially overlap with centrin (Figure 3 Supplementary 2C). At this stage, PLK4, the master regulatory kinase, and SAS6, one of the first centriolar components are either absent or present as small foci within the cloud, often on the wall of the parent centrioles (Figure 3B-C)." some arrows to highlight this would be useful - difficult to see?

We have tried to make arrows on what is now Fig. 3 Supplementary 1 G, but there is to many CENTRIN colocalizing with PCNT. We have enhanced the contrast of the merge to make it more visible.

(6) Figure 3I legend - what are the arrows pointing at? Yellow and white on inserts? ". Around the same time as tubulin, centrin is also recruited to procentrioles (Figure 3I). This stage is probably the stage that we previously documented as A"

However you see centrin at DEUP1 foci in D, and you don't show any eg. SAS6 or PLK4 positive DEUP1+ structures lacking centrin specifically, centrin seems to be present on all the procentrioles in Figure 3I. Did I miss it where you show centrin negative procentrioles in the cloud?

Fig. 3I (now Fig. Supplementary 1J), yellow arrows are pointing at centrioles with non-acetylated MT while white arrows point at acetylated MT. This is now indicated in the legend.

Regarding CENTRIN, it is present as a diffuse staining around the centrosome since the very beginning of amplification (now in Fig. 3 Supplementary 1A with different contrasts), in addition to compose the parental centrioles. This staining can therefore overlap with DEUP1 staining when DEUP1 appears (Fig. 3 Supplementary 1B, E) but not necessarily. In live we observe that CENTRIN and DEUP1 foci can move independently at early stages (Fig. 2 Supplementary 1B, video 2). This is later on, as shown now in Fig. 3 Supplementary 1J (previously Fig. 3I), that procentrioles are all strongly positive for CENTRIN.

A new paper (Laporte et al., Cell 2024) recently showed that the recruitment of CENTRIN on duplicating procentrioles first occurs at the distal end, visible by a small dot, and then appears gradually at the level of the inner scaffold when procentriole reach 160nm, the stage where POC5 appears, which corresponds to the A-to-G transition in our MCC progenitors (Al Jord et al., 2014). One can therefore consider that the same is happening in our cells, and that, with the CENTRIN cloud, we have difficulties to detect the distal CENTRIN dot. We have changed the text to add this reference and discuss CENTRIN apparition in MCC procentrioles.

(7) " The DEUP1 asymmetry previously described at the centrosomal daughter centriole (Al Jord etal., 2014) becomes visible in some cells during the cloud stage (Figure 3B, N; Figure 3 Supplementary 2B) and in a majority of cells" difficult to see - maybe enlarge and single channel from Figure 3F-H in the supplemental Figure 3 to emphasise this?

We have either changed the pictures or the contrast to be more representative with the quantifications. This is visible in Fig. 3A, D, E, G; Fig3. Supplementary 1E and now using correlative light and EM in Fig. 2 Supplementary 2, 3, 4 and Fig. 4 Supplementary 3-4. One has to look carefully at the daughter centriole (marked “dc”). We have not zoomed in since previous manuscript have already described this at later stages with bigger deuterosomes. You can refer to main or supplementary figures in previous manuscripts (Al Jord 2014, Khoury Damaa 2024) where serial sections span the entire deuterosomes and daughter centrioles and show, with nanometric resolution, that both strutures are frequently sticked to each others on tens of nanometers.

(8) Do you have videos of DEUP1 oscillations with nocodazole to show a lack of oscillations?

We have now added videos of DEUP1 oscillations under nocodazole and dynapyrazole treatments.

(9) "In addition, co-staining of centrioles and nuclear pore proteins show a tight colocalization(Figure 6 Supplementary 1B)." I see the colocalisation in panel 1 but less obvious with panel 2 maybe have some more zoomed in panels and some quantification of the colocalization? Is it more striking at the G stage than the D stage?

We edit and change the phrasing as ‘colocalization with NPC’ is not the good term. There is too many centrioles and NPC, they cannot do otherwise than colocalize… What we want to say is that there is a tight connexion with the nuclear envelope. This is why we present a single z, to show that centrioles and NPC are on the same z-plane. This is also clearly visible for centrioles that are loaded on deuterosomes that are around the nuclear membrane in the XY plane. We also added a video to show an entire z-stack of this kind of staining.

(10) "Indeed, SAS6 normally disappears from procentrioles when centrioles are docked, just beforeciliation (Al Jord et al., 2014). This suggests that centrioles were able to degrade SAS6, a process also dependent on APC/C (Strnad et al., 2007), but failed to disengage from deuterosomes." Figure 6 Supplement 1E-F - are you sure it wasn't that Sas6 wasn't loaded correctly at the earlier stage and so is reduced recruitment rather than premature disengagement of Sas6? If it is indeed premature disengagement of Sas-6 - what about CP110 - does the CP110 get loaded and is it still present in noco treated cells arrested in the D phase?

We do not observe SAS6-negative procentrioles on deuterosomes at G-stage but only on deuterosomes in D-stage cells (cells with partly disengaged procentrioles). This is why we hypothesize that, because of the long duration of D-stage and knowing that SAS6 is finally degraded at the end of amplification (Al Jord et al., 2014), we are in the presence of cells where SAS6 has been degraded but where centrioles did not manage to disengage. This is now clarified in the text.

(11) Can you track deuterostome splitting live? Maybe not enough spatial or time resolution?

One has to monitor in 3D (multiple z because deuterosomes move a lot), 2 colors, high temporal resolution (dt=2-5’; to be able to track a single deuterosome), and long duration (deuterosomes are sometimes touching each other and then moving away, giving the impression that they split). This eventually leads to the bleaching of the mRuby fusion protein… We have put an example of what we think is a deuterosome splitting in Fig. 6E (former Fig. 7D). But we decided to finally monitor with low temporal resolution (dt=40’) to avoid photobleaching, and analyze numerous deuterosomes and cells to quantify the number and size of deuterosomes over time in single cells.

(12) The MT nodes - can you segment the tyrosinated MTs and define nodes and then quantify theDEUP1 presence on them?

Please see answer to reviewer 1 regarding this point.

(13) Figure 8 supp 1 (E): Representative XY distribution of CEN2-GFP+ centrioles at the end of migration (Sas6 negative) in brain MCCs treated with DMSO, Nocodazole 1µM and 5µM (48h). Scale bar, 5µm Bit more detail on how you define fully migrated vs still migrating centrioles in z. You say you are using Sas-6 negativity to define fully migrated cells in the legend, yet you say noco treatment leads to premature sas-6 negativity, and yet the apical migration takes longer upon noco treatment?

Nocodazole does not lead to premature SAS6 negativity but to a partial disengagement which lead to SAS6 negative “mature” centrioles being still connected to deuterosomes. We define complete migration when all the centrioles are on the apical side of the nucleus. We now clearly define what “apical” migration stands for in the main text and changed the pictograms in Fig. 8G to clarify this.

(14) Figure 8H and video 18 - it isn't obviously clear to me that the noco-treated cells are "more erratic" or how you decide what counts as apically migrated successfully. How do you control for drift in z? Can you track individual centrioles as you did in untreated and define what is "erratic about their movement?

Erratic means that the centrioles are moving away from each others, and back, in a non-predictable way, instead of migrating up and gathering. The drift in z of the whole cell is visible because there is always some centrioles, that are apically located at the beginning, that remains on the apical membrane, probably because they are already docked.

We have indeed followed the centrioles individually in the nocodazole condition. However, in the control, the XYZ coordinates of one of the centrioles of the centrosome, which normally don’t move, are substracted to the coordinates of all the other centrioles as explained in the method section. This allows to have a subcellular reference, and to circumvent the movements of the cell, which are non-negligible at all at this timescale. In the nocodazole treated cells, the centrosomal centrioles share the erratic movements of the other centrioles and can migrate up and down, which exclude them as a reference. Since the nucleus is also moving a lot, we were left with no reference point.

(15) Figure 8 supplement 1E can you quantify the final area of centriole patch in XY upon noco treatment?

It was in main Fig. 8J and is now in Fig. 8 Supplementary 1F.

(16) Figure 8J legend- MBB is never defined as an acronym.

Thank you for pointing this.

(17) Define what is the frequency and how is it calculated - Figure 8J.

This is the MBB patch area in µm²

Text edits:

(1) "Altogether, these results suggest that, in this non-tissue-specific proxy of MCC progenitors, microtubules organize the onset of centriole amplification in the pericentrosomal region."

Sentences have changed.

(2) "Increasing the temporal resolution to 5-15s reveals that DEUP1+ foci observe an exhibit oscillatory dynamics to at the centrosome (Figure 2E, colored arrows, Video 3, 5/10 cells observed for 1-4min)."

Sentences have changed.

(3) "stage procentrioles were involved in this perinuclear migration and distribution. In fact, this dynamic is reminiscent of the centrosome migration that occurs during the G2-to-M progression in cycling cells in preparation for mitotic spindle organization. In cycling cells, this" Grammar - maybe change to "stage procentrioles were involved in this perinuclear migration and distribution. This is reminiscent of the centrosome migration that occurs during the G2-to-M".

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(4) "We then wondered whether these microtubule-dependent dynamics was were required for an efficient subsequent centriole disengagement during the following D-stage."

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(5) "Then, monitoring tens of disengagement movies, we identified a transient stage during which disengaging procentrioles redistribute isotropically in the 3 dimensions, along the nuclear membrane (Figure 6A, 4:30, Video 7) before losing its contact to migrate to the apical surface (Figure 6A, 6:30 to 14:00)."

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(6) Discussion: "Since pioneer electron microscopy studies on basal body production in quail oviduct MCC 35 years ago (Boisvieux-Ulrich et al., 1987, 1990; Boisvieux-Ulrich et al., 1989), this work is the first to assess the role of microtubules in the now finely described centriole amplification process. This"

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(7) "Using live imaging on brain MCC, we highlight the existence of a nest composed of DEUP1, PCNT and Centrin2, pre-assembled before the onset of centriole amplification onset."

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(8) "Recently, formation of DEUP1 pure condensates in solution as well as FRAP experiments after overexpression of DEUP1 in MCC progenitors suggested that deuterosomes where are not liquidlike structures (Yamamoto & Kitagawa, 2019). Consistently, we never observed fusion events of DEUP1."

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(9) "This reminds is reminiscent of the centriole-to-centrosome conversion occurring at the G2-M transition followed by the associated microtubule dependent nuclear migration of new centrosomes at mitosis onset (Agircan et al., 2014)."

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(10) "Following individual trajectories requires high resolutive resolution spatio-temporal live imaging while avoiding excessive light exposure which disturbs centriole migration (Boudjema et al., 2024)."

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(11) "Using high temporal resolution microscopy, we further identify that individual dynamics is are complex and can be splitted between divided into the baso-apical migration, where centrioles move in a processive and more..."

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Reviewer #3 (Recommendations For The Authors):

(1) Growing MEF-MCCs on micropatterns has successfully mimicked the dynamics of centriole amplification in brain MCCs, allowing the authors to study the spatial origin of procentrioles. Since this is a powerful system, a more quantitative description of the system will be informative and beneficial for future studies. For example: What is the efficiency of this system? Do the cilia that form in MEF-MCCs motile?

The system of MEF-MCCs has been described in a previous paper from the Kintner lab. It seems that growing the MEF-MCCs on micropatterns did not ameliorate the ciliation which is partial, probably due to the absence of an apico-basal polarity.

(2) Figure 2: The analogy drawn by the authors between DEUP1 oscillatory dynamics and centriolar satellites is intriguing. In early amplifying cells within the cloud, do these DEUP1 structures co-localize with the satellite marker PCM1?

We have added immuno stainings of PCM1 in mRuby-DEUP1 / CEN2-GFP cells in Fig. Supplementary 2E. Within the centrosomal cloud, DEUP1 colocalizes with PCM1. Interestingly, this PCM1 concentration at the centrosome is dependent, at least in part, on dyneins. Then, PCM1 can localize around the deuterosomes, but it is never colocalized with deuterosomes (not shown). This is also showed by immuno-EM in Zhao et al., 2019. Although it was shown that PCM1 is a proximity interactor of DEUP1 (called ccdc67 at that time) by Firat-Karalar et al., 2014., absence of PCM1 staining on deuterosomes does not favor the hypothesis of PCM1 and DEUP1 being part of the same entities. One could hypothesizes that DEUP1 is transcribed locally within the satellites, explaining the colocalization of the 2 proteins and the + BioID results, and then form PCM1negative deuterosomes.

(3) The authors propose a physical link between deuterosomes and centrosomes based on their oscillatory behavior. How are the oscillatory dynamics of DEUP1 affected by nocodazole treatment or inhibition of microtubule motors (i.e ciliobrevin treatment)?

These oscillations are inhibited by nocodazole (Fig. 4D). They are also inhibited by dynapyrazole (Fig. 4D). We never succeeded in having a nice disruption of the Golgi apparatus with ciliobrevin and therefore we did not used it.

(4) In addition to nocodazole treatment, it would be important to determine the consequences of microtubule stabilization by taxol and inhibition of microtubule motors during critical stages of centriole amplification where microtubules are reported to play a role for the first time in this manuscript. Another interesting area of investigation will be to study the extent to which microtubule PTMs contribute to these processes.

We now blocks dyneins during the different stages of amplification. The results are in main and associated Fig. 4, 5, 7, 8. The role of microtubule PTM, is not in the scope of this manuscript.

(5) Describing microtubule dynamics along with Centrin/DEUP1 dynamics will be informative in assessing whether these structures associate and/or move along microtubules? Have the authors performed their imaging experiments with SIR tubulin?

Yes, we have tried hard! But we have encountered different obstacles:

3-color video microscopy is phototoxic,

siRTubulin is bleaching very rapidly

The density of microtubules in MCC makes the observation hardly informative

(6) Figure 5: The role of PLK1 in centriole-centrosome conversion and generation of multiple MTOCs can be tested with a PLK1 inhibitor for further confirmation.

We have also tried but inhibiting Plk1 blocks the A-to-G and G-to-D transitions so it was not possible to uncouple the role of Plk1 in stage transitions versus centriole maturation.

(7) Figure 6: The tight co-localization of nuclear pore proteins with centrioles poses questions about the role of nuclear pore proteins or other nuclear proteins that are associated with centrioles during centriole disengagement and migration. Considering the existing literature on centrosome-nucleus attachments, can there be a way to test this question within the scope of this manuscript?

We have tried to deplete Nup133 but it’s killing the cells. Our additional experiments now show that the nuclear migration of centrioles during G-stage is dynein dependent, reinforcing the parallel with centrosome migration in prophase. We also added results from our scRNA sequencing (Fig. 5 Supplementary 1) showing that some key players of centriole migration to the nuclear membrane are conserved in the MCC cell cycle variant, and expressed with a comparable dynamics as to the canonical cell cycle.

(8) Figure 8: Manually tracking a subset of migrating centrioles to define their dynamics during centriole migration and docking provides valuable analysis for determining the molecular mechanism of these processes. In addition to microtubules, does actin contribute to this process? Since centrioles eventually migrate to the apical side in nocodazole-treated cells, there should be other molecular players involved in this process.

We did block actin polymerization but we found that the different stages were affected and that it would be better to dedicate a whole manuscript on the role of actin during each stage of amplification. We discuss the migration mechanism, and the putative role of actin, in the discussion.

(9) The legends for Supplementary Figures 1 and 2 in Figure 3 are mixed and need correction.

Figures have been remodelled.

(10) In Figure 3P, the term "PLK4+" is labeled in bright green, which is not clearly visible. It maybe beneficial to change the color of this label for better visibility.

We have tried to correct this.

(11) Figure 6F quantifies "% tethered flowers" on the nuclear membrane. When quantifying, is the3D localization of DEUP1 flowers in both DMSO- and Noc-treated cells considered? A flower may appear to be on the nucleus in 2D, but it could be detached from the membrane in a 3D view.

The quantifications are done in 3D. However, flowers that are below or above the nucleus are not quantified since the space is confined and the resolution in z to small to see whether they are connected or not. This is now precised in the legend.

Before the editors proceed with an updated assessment, they've requested that we pass on some of the comments that have arisen as part of the evaluation of your revised manuscript. They feel that these concerns should be addressed before we proceed with issuing a formal assessment and publishing the revised Reviewed Preprint:

We thank the reviewers and the editors for the corrections and insighfull comments. We apologize for our delayed answer and hope our corrections in the main text and some of the figures will give them satisfaction.

The revised manuscript is greatly improved with nice new data regarding the role of microtubules. It also has changed quite a bit including the title. The new focus is on the cell and centriole cycle variants in MCC. While this helped to focus the study, there remains an important issue related to the interpretation of the data and the proposed 2-in-1 cycle model. Before providing the final updated assessment, we ask you to address the following points (which were raised already in the first round of review): The manuscript still contains statements that are not aligned with published work and the current view in the field regarding the timing of events during canonical centriole biogenesis. These timings are in conflict with your model that 2 centriole cycles are "superposed" in the MCC cell cycle variant, as currently presented. An alternative straightforward interpretation would be that multiciliogenesis uses an accelerated centriole duplication cycle where key steps occur concomitantly or in short succession instead of being separated by mitotic divisions as in the canonical cycle.

We do agree with the acceleration of all steps into only one cycle, this is actually what we think we have proposed. When correcting our confusions as regard to centriole-to-centrosome conversion (as explained below) and putting the events in a scheme, this reveals that the events of the two canonical cycles nicely superpose, both in term of molecular composition and dynamics (corrected Fig. 9). We therefore maintain that the null hypothesis is that the acceleration is done through a superposition of events that; although driven by the same molecular machinery, are normally occuring in two consecutive cell cycle. We explain ourself briefly in two paragraphs, before answering point by point to the questions of the reviewers.

As regard to centriole-to-centrosome conversion:

We thank the reviewer for pointing out that we used “MTOC conversion” for what is normally called “centrosome maturation”. We have removed the term “centriole-to-centrosome conversion” during the first round of revision but we now realize that “MTOC conversion” leads to the same misinterpretation as regard to the literature on centriole duplication.

The reviewer asks us to refer to the work of the Tsou lab (Wang 2011, reference now added in the manuscript) showing that daughter centrioles are “modified” (e.g. recruit PCM, become competent for MT nucleation and duplication) during late M/early G1. This “centriole-to-centrosome conversion” can’t occur for our procentrioles at this stage since they are not even born during the mitosis that precedes MCC differentiation. Also, in our cells, such modification does not include the capacity to become competent for duplication since we know that procentrioles become basal bodies without making any round of duplication (Al Jord et al., 2014).

Also, we have not done the experiments to tackle the question on when our centriole become “modified-like”. What we can say is that during A-stage, they become progressively positive for PCM (Fig. 5 Supplementary 2) and a weak signal shows that some MT are seen emerging from them (Fig. 5 and Fig. 5 Supplementary 2, and see point by point answer).

What we do see is that, at the A-to-G transition, they increase their PCM recruitment, show clear and strong MTOC ability (sometimes as strong as the centrosomal centrioles), and that this is associated with migration and separation of centrosome/deuterosomes around the nuclear membrane (Fig. 5). We therefore connect this to what occurs at the G2/M transition which is an increased recruitment of PCM protein, an increased ability to nucleate MT, associated with centrosome migration and separation at the nuclear membrane. Since this process in the canonical cell cycle is called “centrosome maturation”, we therefore should refer to this term in our study. However, centrioles in the MCC variants are not organized in centrosomes, so we now compare what we see to the “centrosome maturation” of the canonical cell cycle with an associated reference (Joukov et al., 2018), but name it “centriole maturation”.

We have modified the text (track changes visibles) and the schemes (Fig. 5, Fig. 5 Supplementary 1 and 2, Fig. 9, Fig. 9 Supplementary S1; new versions uploaded) accordingly.

As regard to 1.5 or 2 cell cycles

Except for the “MTOC conversion” that we have now changed, as explained above, we think our work does suggest (depicted on Fig. 9) what the reviewer states for centriole duplication: “In the current view, centriole biogenesis starts in early S, elongation proceeds through G2/M and by early G1 it is complete. During M/early G1 centrioles disengage and newly formed daughters recruit PCM (centrosome conversion). Then these centrioles go through another complete cell cycle and when they reach early G1 again they have acquired DAs and SDAs. Key here is that biogenesis and disengagement/centrosome conversion are separated by the first mitosis (ensuring duplication occurs only once), and acquisition of DAs and SDAs is separated by another mitosis (ensuring that cells only form a single cilium)”.

We feel that going from early S to a G1 phase, after 2 mitosis, is what one can call “2 cell cycles”. One of the paper that inspired us a lot when studying how the cell cycle machinery can drive centriole amplification in MCC is a paper from Jadranka Loncarek team (Kong et al., 2014) where they also state that “nascent centrioles gradually mature through 2 cell cycles”. Very interestingly, in this study they show that when they enhance Plk1 activation, they could erase centriole age and new procentrioles are able to recruit PCM and appendages within only 1 cell cycle, without mitotic progression, like what we see in MCC. We have added the reference in our discussion.

Point by point answer

(1) Original work on canonical centriole disengagement and centriole-to-centrosome conversion should be cited (e.g. PMID: 16862117, PMID: 21576395)

As explained earlier, we used the wrong term since the begining. We do not speak about the centriole-to-centrosome (nor MTOC) conversion since we do not test when centriole modification (Wang et al., 2011) occurs in the MCC cell cycle variant. We know that PCNT is present on the procentrioles during A-stage (as shown in Fig. 5 Supplementary 2B), but we do not know when it is recruited (UExM did not work properly with this antibody). We quantify a weak MT staining in regrowth experiment during A-stage and see that procentrioles can be connected to MT in both brain MCC and MEFs (as shown in Fig. 5D, E for brain MCC and Fig. 5 Supplementary 2F for MEFs) , but we do not know when during A-stage they become competent for nucleation. We therefore did not speak about this process that we do not document. What we clearly document/quantify is the enhanced MT nucleation capacities at the A-to-G transition, concomitent with the nuclear migration (easily defined with Cen2-GFP or GT335 stainings) and that we compare to centrosome maturation occuring at the canonical G2/M transition.

(2) The authors state in several places that canonical centriole formation and maturation takes two iterations of the canonical cell cycle. This is imprecise. Based on the above work and work by others, the broadly accepted view is that it takes 1.5 cell cycles. This difference matters for the final proposed model (see below). Reviewed e.g. here: PMID: 20869612; PMID: 30601682

Our answer is in the preamble.

(3) "Centriole maturation cycle superposes with centriole elongation cycle in the MCC cell cycle variant": Your description of the canonical cycle differs from the current view in the field. In the current view, centriole biogenesis starts in early S, elongation proceeds through G2/M and by early G1 it is complete. During M/early G1 centrioles disengage and newly formed daughters recruit PCM (centrosome conversion). All this occurs in 0.5 cycles. Then these centrioles go through another complete cell cycle and when they reach early G1 again they have acquired DAs and SDAs (total of 1.5 cell cycles). Key here is that biogenesis and disengagement/centrosome conversion are separated by the first mitosis (ensuring duplication occurs only once), and acquisition of DAs and SDAs is separated by another mitosis (ensuring that cells only form a single cilium).

(4) Fig 5A, B and Fig. 9

(a) Are 2 separate figures needed for the model? They seem redundant.

We find it easier not to wait Fig. 9 to have the first part depicted.

(b) The model shows loss of SAS6 throughout G1, but this already occurs during M/early G1

Thanks. It was already ok in Fig. 9, we have modified for Fig. 5.

The model shows "MTOC capacity/conversion" during S phase, but this occurs during early G1

Thanks a lot, as explained earlier, we used the term MTOC conversion occurring in G1 for what is normally called centrosome maturation occurring in G2/M, as explained earlier. We do not speak anymore of MTOC conversion since we have not tackled this question (explained above). We have therefore removed MTOC conversion in the texts and the schemes and replaced it by “centrosome maturation” for the duplication cycle, and by “enhanced MT nucleation capacity” for the MCC cycle. To be clearer and schematize that procentrioles are competent for MT nucleation before G2/M or A/G transitions, we have added some MT nucleated from G1 procentrioles during the canonical cycle, and from late A-stage procentrioles during the MCC cycle.

The model shows disengagement only in the second M phase, but this occurs already at the first M phase, directly following centriole biogenesis, right before centosome conversion.

This is a big edition error in both Fig. 5 and 9. Of course the daughter centriole disengage during the first M-phase. This has been changed. Thanks a lot for spotting it. This, however does not contradict the hypothesis of superposition.

We also added the acquisition of distal appendage which was written in Fig. 5 but not in Fig.

9 for duplication during the second M-phase.

When the correct timings are incorporated in the figure, the proposed superposition of two cycles is not an accurate description of the events. Instead, your data seem consistent with a model where MCC incorporates all steps in one cell cycle variant that lacks mitoses, so that disengagement and MTOC conversion occur together with centriole elongation, followed immediately by acquisition of DAs and SDAs.

We do agree with the acceleration of all steps into only one cycle, this is actually what we tried to propose. When putting the events in a scheme, this reveals that the events of the two canonical cycles nicely superpose, both in term of molecular composition and dynamics (Fig. 9). We therefore maintain that the null hypothesis is that the acceleration is done through a super opposition of events that; although driven by the same molecular machinery, are normally occurring in two consecutive cell cycle. This is notably consistent with the findings of Kong et al., 2014 cited previously.

(5) While all reviewers felt that there was no need to introduce the new term "nest", they leave it to the authors to keep it. However, the authors may want to consider that the term is still not introduced and explained properly, which may confuse readers. For example, while this section reads like an introduction to the term: "Correlative DEUP1 live-imaging and EM highlights the existence of a pericentrosomal "nest" in brain MCC", the term is already used two times before without explanation. The first mentioning is at the beginning of the results section and is followed by citations, which gives the impression that these studies describe the nest, which is not the case.

The first mention of “nest” is in the end of introduction resuming the findings of the paper where the term is in the following context: “we found that centriole amplification emerges in a pericentrosomal “nest” concentrating core centriole/deuterosome elements”. We looked at nest definition in the Collins Dictionnary : “a structure or other place where creatures, esp. birds, give birth or leave their eggs to develop”, we felt this was clear. We added quotation marks around the term nest.

Then, the result section opens with this sentence: “The origin of amplified centrioles in MCC remains controversial. Some live imaging experiments and electron microscopy suggest that the centrosome could constitute a nest for centriole and deuterosome biogenesis (Al Jord et al., 2014; Kalnins et al., 1972; Mori et al., 2017), but others have proposed that procentriole-loaded deuterosomes emerge independently from the centrosome location, all over the cytoplasm (Nanjundappa et al., 2019; Sorokin, 1968; Zhao et al., 2013, 2019).”. Here, the term nest is again used as a place of birth for centrioles and deuterosomes which is what is actually proposed in these papers. First, Kalnins el al., in 1969 (we made an error on the reference date, this has been changed), resume in their abstract “This observation suggests that all of the clusters may form initially in close association with the diplosomal centrioles”. Then, not to mention Al Jord 2014 which comes from our lab, the title of Mori et al. is “Cytoplasmic E2f4 forms organizing centres for initiation of centriole amplification during multiciliogenesis”, and in the paper, they show that E2F4 accumulates at the centrosome. This is now also proposed by collaborators for MCIDAS (Lu et al., 2025). We feel that these references, which are often omitted, are appropriated at this location.

Then we continue with: “To test whether microtubules drive the organization of a centrosomal nest from which procentrioles emerge”, which keeps the notion of the place of birth.

Then the title "Correlative DEUP1 live-imaging and EM highlights the existence of a pericentrosomal "nest" in brain MCC" arrives. In this section we first speak about a pericentriosomal cloud on which we zoom in using CLEM, to then conclude at the end of the section “Altogether live imaging mRuby-DEUP1/CEN2-GFP during early A-stage suggests that core deuterosome and centriole components are concentrated in a primordial cloud around the centrosome, which constitutes a nest where centrioles and deuterosomes concomitantly form before they move away from the centrosomal region (Fig. 2F)”.

Finally, we begin the discussion section regarding the nest by: “We named this transitory compartment a “nest” since deuterosomes and procentrioles emerge specifically in this region and grow while moving away from it.”

During the first revision, we tried to make it clearer. If this is still not the case after and the reviewer has another proposition of definitions/phrasing, we will be glad to consider it.

As replied to the other reviewer, the term “nest” does not need to be retained as a new terminology. It is just a way for us to identify the transitory region and to best define one of its function/characteristic which is to host the birth of new deuterosomes and centrioles.

The following comments from Reviewer #3 may also provide further context regarding the editors' remaining concerns:

The authors have done an excellent job addressing the points I raised overall, and the revision is substantially improved in focus and clarity. That said, some concerns raised by other reviewers, particularly regarding terminology and statistical analysis, could have been addressed more fully. One issue remains insufficiently resolved. Several quantitative analyses (for example Fig. 5C and 5E) still appear to rely on pooled single-event measurements collected across three independent experiments. This approach can overstate statistical significance. The authors indicate in their rebuttal that they use chi-square tests to compare proportions and to justify pooling across replicates. However, I am not convinced this addresses the issue for the intensity-based and single event distributions shown in the panels specified above. I recommend that these key analyses be represented with biological replicates shown explicitly (superplot-style, with replicates distinguished).

Our reply was for the comparison of proportions and not the intensity-based and single event distributions shown in the panels Fig. 5C and Fig. 5E. We have now changed our plots to represent biological replicates explicitly (superplot-style, with replicates distinguished). As for the statistical analysis: we evaluated differences in marker intensity between A-stage and G-stage samples using a linear regression model, with stages as the main effect and replicate as a fixed covariate, to account for batch variation. Statistical significance was assessed using Type II ANOVA.

Separately, I continue to feel that some newly introduced terminology (for example, the "nest") may not be necessary at this stage. It may be sufficient to describe these structures and focus on their spatiotemporal behavior, composition, and measurable features, rather than assigning new names. Having read the authors' response, I understand that they would like to retain this terminology, which is acceptable; however, it may not be readily adopted by the field.

The term “nest” does not need to be retained as a new terminology. It is just a way for us to identify the region and to best define one of its function/characteristic which is to host the birth of new deuterosomes and centrioles.

Minor correction (remove "in MCCs" part from the following sentence):

In MCC, PCM1 depletion alters deuterosome formation and centriole production in brain and airway MCC (Hall et al., 2023; Zhao et al., 2021).

Done

This reminds me of how the 2019 elections in India were won by using WhatsApp. The winning political party had a network of unofficial party workers present in informal community WhatsApp groups at the district and sub-district levels. The messages to be sent to these groups across India were dictated by a centralized IT cell - messages mostly containing misinformation and spreading hate (Refer to this article - https://pulitzercenter.org/stories/inside-bjps-whatsapp-machine). I shudder to imagine what would happen if these people in power were applying similar strategies through AI - according to the digital potemic villages example in China, this is already underway. I also feel that there needs to be a certain already existing orientation for one to want to consume certain content. Referring to the sycophantic nature of AI, this implies that if someone is already inclined to be a racist, etc., the AI is going to reinforce this orientation.

This is very interesting for me, and it reminds me of Julia Cameron's work on Language, Sexism and Misogyny. There was a chapter on language policing which talks about how negative judgments about how a group of people speak is often correlated to prejudices one may have about a group of people. Sometimes, it also leads to us hearing something that they might not have objectively said. This also leads to the people who are considered 'others' policing their own language.

Gramsci’s concept of hegemony includes schools, newspapers and other social structures working together to enforce capitalist ideals, creating a society where these ideals are considered to be ‘common sense’. This reminds me of the three logics in contemporary society, discussed by Dr. Stewart on Friday morning. Gramsci’s idea of cultural hegemony in today’s world feels closely linked to the overwhelming influence (power?) of the logic of business over both media and education. The logic of business in contemporary society feels culturally hegemonic, and the use of big AI in schools without sufficient guidance or policy could further cement the existing inequities in our society between these three logics.

I once asked my supervisor how come it takes so long to create change? This was about a month ago. He explained that all these inequities and hierarchies are created and rooted in the foundations of structures, and to change it, we must break down these structures. But breaking down these structures take time because there are always people who benefit from them that want to keep them in place. This sentence reminds me of how normalized inequality and colonialism is that sometimes we don't notice, but it's just considered 'common sense'

This reminds me of what someone in the class said to me. They expressed how easy it is for misinformation to travel, but not only travel, but how difficult it is to correct misinformation with factual information. When people such as Elon use their personal biases and opinions to spread misinformation on such large platforms, it ultimately removes the sense of agency. Also, when trying to correct misinformation, most of the time the factual information does not get as many views compared to the misinformation

this reminds me of the fact that the transform of a society may not necessarily result from a radical revolution or a war, and it is the transformation of people's minds being the key.

I appreciate that the author includes this because it pushes back against the idea that every historical figure was trying to start a larger movement. Sometimes people are simply trying to protect their own family. Looking back, it's easy to expect every legal challenge to represent a broader fight for justice, but this chapter reminds me that many of those cases started with one parent trying to do what they believed was best for their child.

Edoardo Foschi conversation, is that who this reminds me to ping?

I love this idea because it made me think more deeply. It reminds me of a seed. A seed already has the potential to become a particular kind of tree, but it needs many different factors such as water, sunlight, nutrients, and time to continue growing. Eventually, it becomes a strong tree that benefits the world by supporting the ecosystem and providing shade for others.

I think human development is similar. We don't suddenly gain completely new abilities as adults. Instead, we build on skills that already exist in an early form during childhood. For example, when we are five years old, we learn to tie our shoelaces. Later, we learn how to solve problems in school, such as passing an English test. As adults, we may solve much more complex problems, like running a business or leading a team. The skill is still problem-solving, it has simply become more advanced over time. That is why I love the idea of continuous development.

Las Casas' description of the Native people as innocent sheep not only negates their ability to be seen as individuals with their own complex lives, but it also reminds me of the saying "to be led like sheep to the slaughter", insinuating that what the Spaniards inflicted upon them was brought on simply by the fact that they are "sheep". La Casas even further exploits this idea by implying that the slaughter is one of God's will, "qualified by the Almighty". While Las Casas may not agree with the exploitation of the Native people; he is a priest and uses God as an excuse for ill-willed intentions, which is rather blasphemous.

Reminds me of an experiment summarized in Barry Schwartz's Paradox of Choice. As I recall it, grocery store customers were divided into 2 groups - one group saw shelves upon shelves of different jams and jellies; the other group saw about 4 kinds of jelly. Everyone agreed that the store with more options must be the better store. But the people presented with fewer options said they had a better shopping experience.

And they bought more jelly.

Reminds me of 2 posts by Maya C. Popa: Want to write better poems? Stop sharing them so quickly. and [Three poems I cut from my next book] (https://mayacpopa.substack.com/p/three-poems-i-cut-from-my-next-book). I say this as someone who's not afraid of "first thought best thought" creativity, but that does need to be balanced by thinking realistically about one's goals.

This reminded me of the nail video from class and also reminds me of a phrase I saw online. "Do you want solutions right now or do you just want me to listen?" It was from someone making a joke/complaint about how aren't able to use context clues and emotions to draw accurate conclusions about their partner's needs. This was the solution given by the creater of the video. While the phrase itself is an attempt at trying to understand a situation better. I can’t help but image hearing this in the middle of an emotional moment would make me feel a bit disconnected from my partner. In my mind it implies that they don't have good listening skills or the ability to read any context clues.

This reminds me of seeing how differently students respond to bilingual teachers in dual immersion classrooms or classrooms with many English learners. And how it is imperative to have some bilingual teachers at least on staff for support and guidance both for language development and cultural understanding and reverence.

I asked myself why they were white fathers, and quickly recalled that men were the only people with social agency at the time. Interesting to consider how any men at the time joined the tribe. It reminds me of how in many cultures, mothers are considered responsible for how we determine if someone is of an ethnicity or educated on their ancestry and cultural practices. And, as teachers are so frequently connected to the role of a parent, how it is our responsibility to be educated and considerate of the cultural practices and beliefs of our students to be effective & respectful members in our academic community.

This reminds me of how missionary schools treated (and mistreated) indigenous children. All correspond with the purpose of schools to create good (anglo-americanized, Christian, conservative) citizens.

It reminds me of how we have seen a pendulum swing from trade schools being considered appropriate to rarely discussed in schools as an appropriate life/career path - with an over emphasis on higher education. And I hope, as educators we ensure students feel all paths are valid and available to them. And how when there are arguments about how public education is to "just make workers for the workforce," we can argue for quality education while not being dismissive about the intelligence and integrity of blue collar workers.

This reminds me of how many of us simultaneously grapple internally (and at times externally) when we hear, "liberty and justice for all," and many similar encompassing phrases, knowing they are referring to one group. And how sometimes our job feels like it is equal time innovating new strategies and questioning and dismantling oppressive ones.

When I read I normally look through it with a slightly pessimistic or just an analytical view but after reading this I was slightly pleasantly surprised although this woman had some things going against her and experienced many tragedies she still kept not just her faith but also herself in a way. It reminds me of family members that I know that experience similar things and yet still keep their own faith.

This stood out to me because Harjo is saying that Native voices matter and deserve to be heard. It reminds me of my project because people in rural Alaska often do not get much attention when talking about education problems, even though those issues affect real communities.

I find it fascinating how a person's mind will seek out any sort of confirmation even if it comes from ones self. This reminds me of a self-fulfilling prophecy from an earlier chapter. How our behavior and actions will feed into a what we believe. Even to the point of causing a self-destructive cycle if gone unchecked. It also reminds me another chapter where coping mechanisms for stress before giving a speech was covered. I'm sure the breathing exercise for anxiety or stress could also help lessen irritation and there for help get rid of negative nonverbal actions/cluster.

This reminds me a lot of a concept I heard, I believe from a video, a long time ago. It was about grooming habits between people and how it can be a reflection of comfort with physical intimacy. The video talked about the moment in a friendship or relationship when we feel comfortable enough to reach out and 'groom' the other person. That the closer a connection the more comfortable and willing we are to 'groom' the other. Someone in a newer relationship might point out something like a messy hair, lint, or a tag sticking out. As you move up levels of intimacy that moves onto just letting them know you're reaching to fix it. Finally reaching out and fixing the problem you noticed without having to say anything to them. It was supposed to be a sign to see how physically close two people were. As the person being groomed had the chance to reject the help offered and set a boundary. The person asking had a chance to express care for others appearance and or comfort. I'm not sure entirely how true observation was but this section reminded me of the video as it discusses levels of physical contact in comparison to the depth of the relationship.

This reminds me of the previous sections where the text mentioned that some basic nonverbal communication systems are universal. Things like shaking your head no, nodding yes, hands pressed together in thanks, are generally universally known. Same as some of the emblems like a thumbs up, okay, or waving come/go away. It makes me wonder just how far a totally nonverbal interaction could go for people that don't speak the same language. From my experience helping customers that dent speak English at times it was easier to stop trying to translate. Instead focus on the context clues, nonverbal cues, and whatever small words I could recognize.

Reminds me of crime investigators.

This reminds me of that Charlie Kirk quote "When people stop talking that's when violence happens".

This is super important! It reminds me of what Stó:lō writer Lee Maracle says that Canadians often talk about Indigenous peoples rather than to them or with them. How can we talk with people, be in conversation with each other, and not be a container for knowledge?

This reminds me of the term "research creation" that has been introduced to me in this class. What is the distinction between art and scholarship? Relatedly, what should art do?

This reminds me of using emphasis or fluctuations in voice when speaking. Just like a well-placed change of tone gestures can help drive a point across or draw attention to the speaker. I would also assume using both of these together could be a really good way to reengage an audience, show enthusiasm, or make a statement/fact feel more impactful.

This is a very interesting point I haven't considered before. It reminds me of a previous chapter. Going over demographic, environment, and how important it is to consider the audience for a speech. Something like this could be researched before giving a speech abroad or to a foreign audience.

This portion of the chapter reminds me a lot of different ways I communicate with my friends over text. Especially when it comes to our use of images and emojis. It seems like a simple silly thing, but we use emojis and images to help express the tone or emotion behind a text. Adding some sort of visual media along with text is the easiest way for us to give more detail without having a verbal conversation. This can get even more complicated as some emojis we have assigned meaning that has more to do with our personal relationship and less the emoji itself.

This reminds me of what someone in Michael Pollan's book said about how our consciousness is like a stage and what our consciousness decides to put a spotlight on depends on our subconscious.

This reminds me of how people can have thoughts or reactions without actually understanding where they came from. We experience the result but not the process.

Black truffle taste described as nutty and earthy. This reminds me of how people that work at cheese stores tell me how certain cheeses taste.

This section reminds me a lot of what I have seen some people do in arguments. They will acknowledge that the other person reality is different but then insist that it also means its incorrect. I feel like a large part of disagreements are spent on the back and forth of trying to get someone to not just understand but ultimately subscribe one sides version of reality.

I wonder how quickly into a first impression someone assigns a stereotype onto another person. It also makes me wonder if people less exposed to a variety of cultures rely on stereotypes more heavily. I could definitely see someone stereotyping another person based off a first impression just because they only things about that stranger are features or traits that belong to a stereotype. Which reminds me a lot of the pattern searching and connections that was mentioned earlier in the chapter.

I never stopped to think what the first words might have actually been or sounded like. It reminds me a lot of talking with babies or toddlers. How sometimes they assign sounds or gestures to the things they ask for the most. I have even seen kids use a word that sound similar then pair it with hand gestures until someone understands. While it may be confusing to a stranger being around a baby or kid long enough you can start to see the connections and communicate with them easier.

I like this part because it reminds me that the class was not just about finding one “correct” opinion on social media issues. It was more about learning how to think carefully about tradeoffs and consequences. I think that made the combination of ethics and programming especially useful.

This reminds me of the "everything apps" that are quite popular in East Asia. These are apps where many different activities- from navigation to shopping and etc can be done on one app. Of course, there is a convivence to this, but I do worry that it leads to uncompetitive behavior as the companies running the everything apps will curate the experience to favor themselves.

大多数人认为AI应用应该优先采用最新技术快速实现，但作者将其比作云计算早期的'简单迁移'模式，认为这是一种可能导致资源浪费的短视行为。这与当前AI领域的快速采用主流观点相悖，暗示企业在AI应用上可能需要更加谨慎的基础架构规划。

I agree with the opinion that parents should be kind to their children. It kind of reminds me of the trend that was going around on tiktok where parents would crack eggs on their kids head then post them crying afterwards. There was a lot of backlash for that one too, and I think its agreeable that parents should never be making their kids cry on purpose.

good summary, good visuals. underrated video!<br /> the magnetic pole shift situation reminds me of the movie "dont look up".<br /> but obviously, a magnetic pole shift is much slower than an asteroid impact...

This kind of reminds me of one of the Ethics Frameworks that I have reviewed over time; Egoism. Egoism focuses on your own pleasure. An individual with Egoism Lifestyle may do things selfishly and in order to please only themselves.

This concept specifically reminds me of a certain era of YouTube culture where a small number of creators did this regularly. The one that sticks out in my mind is the channel LeafyIsHere. This channel would react to other creators' content and make fun of it. It had several million subscribers, so the account being targeted would receive massive amounts of hate. This channel was widely considered to be toxic as of 2017 or 2018, and is now inactive. It's interesting to reflect on how many people that account was able to influence, and how much negativity was spread.

Reminds me of working meetings with LPMA or at work

This distinction between private and public harassment is important because the harm shifts when there’s an audience. Public attacks don’t just hurt the target; they also shape how other people see them, and they implicitly recruit bystanders as witnesses or even participants. It reminds me of times I’ve seen posts on social media where the real damage wasn’t just the words, but the fact that hundreds/thousands of others were liking and sharing them.

Reminds me of the Garden of Forking Paths by Borges. Also, this is an attitude that might help completionists and perfectionists look at their obsession with "finishing things" in a different way – you can never truly complete anything, only arrive at it in a way different from how you arrived at it the first time. As Venkatesh Rao says in this Ribbonfarms article (https://ribbonfarm.com/2016/05/26/how-to-take-your-brain-off-road/), some pieces of reading are so alien that the first time you read them, you are only learning how to read them. Also explains why good works, like classics, reward rereading more than reading. You never arrive at a classic the same way twice. Like the saying "No man steps into the same river twice."

These bot hate farms are really intersting. It really reminds me of that star wars filmed that had like over 50% of the hate online originate from Russian bots. What are the incentives behind it? I think especially for that star wars one, it really is interesting. Especially as the public perception of that film is AWFUL, but how much of that is real opinion and how much of that is botting? Was the film so bad? I didn't really enjoy it, but it somewhat didnt deserve ALL the hate that it got. I wonder in the future how much bots will have an influence on public opinion- and what the implications of it are. And also like, how much of the internet today (like youtube comment sections) are real people with real opinions?

This reminds me of a lot of true crime videos that I have watched where many of the cases were actually cracked by people on the internet. One that I can think of actually has a Netflix documentary called Don't F With Cats, where the internet identified and caught a loser who was killing cats on video and uploading them.

I think this is a good example of people getting together to help other people. This reminds me of a documentary, "Don't f*** with cats" which is where there was a guy on the internet publishing disturbing content of him torturing cats and people tracked him in order to bring him to justice. This is an example of crowdsourcing.

This article about how a woman had no wikipedia page until she was associated with a male is so sad to me. She literally won a Nobel Peace Prize and is not recognized onliine for it in any public matter. It is only until a man is beside her name that they rcognize it. This is extremley disappointing because id think that an accomplishment that big wouldn't go unrecognized because of gender.

I thought this part was interesting because it shows that moderation is not only about ethics or safety, but also about making a platform usable at all. A lot of arguments about moderation focus on free speech, but this section reminds me that if everything is allowed, the site can become full of spam, scams, or repetitive content and stop being useful to regular users. That makes moderation feel less like an extra feature and more like part of what makes a platform function in the first place.

Though clear and coherent, this conclusion feels like a bit of a let down after such subtle, perceptive, and engaging analysis. I think you might say how, after (or as an extension of ) collapsing the distinction between flaneuse and writer, the poem animates the reader in their combined role, inviting us to experience an autonomous, subjective, mutlisensory interiority, in which the self is not objectified nor objectifies others, but experiences its own power and permeability. In this way, the poem reminds me a bit of Walt Whitman's Song of Myself, which concludes by hailing the reader, saying: if you want me, look under your boot soles. We become Whitman, walking the streets of the newly formed American nation in his shoes.

Theological: This passage reminds me of the Upanishads because the Divine is described as existing throughout all creation. This also reflects the idea that Brahman exists in everything.

This reminds me of what my college coaches used to preach. They also talked about how when you're wearing "our brand" you're representing more than yourself, you're representing our brand and our comminuty.

This reminds me of culture on any team/workplace. You must surround yourself and others with people that are aligned and focused on the same goals.

I looked at KC Green’s post about the “This is Fine” meme. In this post, he explains that the comic became much bigger than he expected. He says the meme is still relatable to many people, but it also feels strange because one old comic became more famous than much of his other work. He also talks about how hard it is for artists to control their work after it becomes popular online. I think this source is useful because it shows the human side of internet memes. People often share memes without thinking about the artist behind them. This post reminds me that memes are not just random pictures online. They are also someone’s creative work. Once a meme spreads widely, it can belong to internet culture, but the original creator may lose control over how people use it.

This reminds me of chain mail I received in the early 2010s via SMS, when I still had a flip phone. Usually there wasn't a monetary aspect of the message. It didn't request that you send money to the sender. Instead it said something like "Send this message to 10 friends for good luck, ignore and receive 10 years of bad luck". I'm really curious what motivated people to create chain mail such as this. It has the make up of a social experiment, but I imagine it would be really difficult to track the movement of the message over time.

I think by doing this could be helpful to try and understand what thoughts were like back then. It reminds me of something in the 1900's saying that in the next century, we will have flying cars, even though we currently don't. It was there point of view and personal belief that humanity would evolve to that point.

This idea emphasizes that teacher should learn from students' environments instead of making assumptions. It reminds me that understanding a students' community can influence how lessons are designed. For example, a teacher might incorporate local resources or address challenges like lack of safe play spaces. This reflects a shift from teaching in isolation to teaching with awareness and empathy.

I found it interesting how dictionaries organize data with keys and values, because it reminds me of how social media profiles have different types of information all in one place. It makes it easier to see how real apps use this structure

I found this source super cool. It talks about a small percentage of women who can see four base colors instead of three. It's super interesting and reminds me of the Psych 101 class I took in the fall where one of the topics we looked at were how do we know if everyone sees the same colors.

This part of the reading stood out to me, the contrast between putting the burden on individuals as well as the designers was interesting. The section on coping strategies made me realize how often people are forced to quietly adapt themselves to systems that were never designed with them in mind like students sitting in front of the classroom without knowing why they struggle to see. That idea felt frustrating because it just normalizes the expectations that individuals should adjust rather than questioning. I personally think the shift toward universal design and ability based design is more ethical and sustainable. It reminds me of discussions in UX design where we talk about designing for edge cases and benefitting for everyone as a whole. I feel that companies should just treat accessibility as a core requirement instead of an extra feature implemented.

On 2022-07-21 14:39:58, user Benjamin Buchmuller wrote:

Very exciting work! Reminds me of Giehr et al. (NAR, 2018) "Hairpin oxidative bisulfite sequencing" https://dx.doi.org/10.1093/... I feel that their work should be cited and discussed in the manuscript.

On 2021-04-23 13:22:31, user Erik Gylfe wrote:

Dear authors,<br /> I find your study provocative and interesting, but somewhat difficult to read due to erroneous reference to Figures. You point out that most previous studies have been performed by stimulating beta cells with unphysiologically high glucose and now demonstrate interesting Ca2+ responses at more reasonable concentrations of the sugar. From this point of view I think you provide important new data using an elegant experimental approach.

However, I am concerned about the experimental design and some conclusions drawn. The data are taken as argument to return to very old and nowadays mostly abandoned ideas that Ca2+ influx has only a minor role during the first minutes of beta cell activation. Some of these old ideas are based on unfortunate experimental design, which I also find in the present study. The common denominator is simultaneously changing two parameters without considering that the timing of<br /> the effects may be different.

I think that the isradipine experiments is a telling example. It is obvious that the effect of the used concentration of isradipine has a slow onset (Fig S3-1) and an even slower off effect (Fig 2) in your system. The timing definitely seems slower than that for the glucose response. Therefore, the most likely explanation of the results is that the Ca2+ channels are not initially blocked and glucose maintains some of its early effects on voltage-dependent Ca2+ influx. The same explanation likely applies to the diazoxide experiment (Fig. S3-2), particularly since the concentration is on the low side. In my experience glucose-induced intracellular Ca2+ release is unlikely when Ca2+ influx is blocked except after artificial elevation of cAMP (see ref 16, which by the way is not correctly cited in the reference list). Instead, the typical<br /> effect of glucose elevation under such conditions is a lowering of Ca2+<br /> due to ER sequestration. However, the latter effect would probably escape detection with the presently used low affinity indicator and is likely rather modest when raising glucose from 6-8 mM.

It is stated in lines 322- 323 “The prominent role for intracellular<br /> Ca2+ release had strong early support from 45Ca2+ flux studies”. 45Ca2+ flux data are often difficult to interpret, and I think you may mix up efflux of 45Ca2+ from islets with intracellular Ca2+ release. Most 45Ca2+ studies are instead consistent with a [Ca2+]i lowering<br /> effect of glucose when voltage-dependent entry is prevented. See for example Bergsten et al. Am. J. Physiol. 255: E422-E427, 1988 for the effect of glucose on both 45Ca2+ and [Ca2+]i in the presence of diazoxide.

The isradipine experiments reminds me about old data taken to indicate that first phase insulin release is independent of Ca2+ influx (Wollheim et al. J. Clin. Invest. 62: 451-458, 1978). In that study first phase insulin release was unaffected when glucose was elevated simultaneously with addition of 5 microM Verapamil. With more effective prevention of Ca2+ influx they might instead have discovered that glucose lowers basal insulin secretion under such conditions (Bergsten et al. 1988).

Although I agree that Ca2+ release from (but also uptake into) the ER are important for shaping the slow [Ca2+]i oscillations, I get the impression that you implicate a more fundamental role of intracellular Ca2+ release in their generation. I think this must be discussed in relation to experiments indicating that the glucose-induced slow oscillations are maintained (with different kinetics) when intracellular Ca2+ uptake and release by the ER is prevented by SERCA inhibition (Liu et al., J Physiol 508: 471-481, 1998; Gilon et al JBC 274: 20197–20205, 1999).

The effects of ryanodine and acetylcholineare interpreted only in terms of Ca2+ release from the ER. I think there is a striking difference in the responses. The effect of acetylcholine is rather similar to that of raising glucose from 6 to 8 mM. This is likely explained by the Na+-dependent depolarizing effect of acetylcholine being sufficient to trigger electrical activity in beta cells exposed to threshold glucose concentrations (see cited ref 22).

The ryanodine data looks rather compelling but are inconsistent with other cited observations. As you acknowledge, the functional significance of ryanodine receptors in beta cells is controversial. I would like to see the effect of repeated exposures to 100 nM ryanodine which is expected to induce reproducible responses since the inhibitory effect of 100 microM ryanodine was reversible.

In this context I lack information about how representative the<br /> observations are between experiments. As I understand it, the illustrations show results from individual experiments. It is stated in Methods “At least 3 slices/mice were used for each experimental condition” but nothing is stated about variation between experiments.

Best regards,<br /> Erik Gylfe

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On 2020-04-23 02:28:24, user David Mayo wrote:

Inspiring, congrats on the great work! This reminds me to what we predicted in our study about prevalence and diversity of type IV CRISPR-Cas systems, which are primarily encoded on plasmids. We observed co-occurrence of certain type IV subtypes/variants with type I subtypes present in the host chromosome (e.g. I-E and I-F). Type IV systems lack adaptation cas casette, and some of them carry cas6e or cas6f. Similarly to you, we hypotesized then that they were sharing the adaptation machinery with the co-occuring I-E/I-F. So we took the most common pair IV-A3/I-E and analyse whether there was conservation in their leader, PAM and repeat sequences, and there was. Moreover the cas6e from IV-A3 and I-E were clustering together, as well as the cas6f from IV-A2 and I-F, providing evidence of common processing mechanism for crRNA maturation.

https://academic.oup.com/na...

On 2019-07-05 22:41:36, user Charles Warden wrote:

Figure 2 in your paper reminds me of Figure 2 in Yizak et al. 2019, except I think that other paper's strategy of showing the range of absolute counts has a more intuitive interpretation than showing residuals (even though I see that you are trying to correct for multiple factors, and you want to show that age affects the differences among the skin-exposed samples)

Was there any communication between the labs? At this point, it might be good to cite that other paper. However, I think that paper could have possibly benefited from waiting a little longer and possibly being in a lower impact journal (and/or being split into separate papers):

http://cdwscience.blogspot....

In general, having similar conclusions from different studies should help provide confidence (although you are both using the same dataset, so it isn't really independent validation). However, I think there can also be value if you take more time to do things carefully (and there can be other factors that delay paper submission). So, I think this is something interesting to think / talk about either way.

On 2019-01-31 19:31:28, user Charles Warden wrote:

While the presented results are different, this kind of reminds me of an earlier paper that I wrote (and have some revised comments / more-public data):

https://journals.plos.org/p...

https://github.com/cwarden4...

In yeast, it seemed like EvoFold predictions in coded regions tended to vary with CAI and were enriched in ribosomal genes (although that is different than your transmembrane emphasis, in a different species).

However, if there is relevance for selection within synonymous sites, maybe some discussion about EvoFold predictions in flys could be useful? I believe I recently had some difficulty finding an executable (if you wanted to define conserved folds in new sequences with EvoFold), but perhaps you could get some data from this paper that used EvoFold predictions in fly:

https://www.nature.com/arti...

(for example, there are also EvoFold predictions in the dm2 UCSC Genome Track)

I see that there was also an "EvoFam" paper (https://genome.cshlp.org/co..., but I haven't tested that program (and I'm also not sure about it's availability). In this case, I don't think it was applied to flys, but perhaps this can help with a literature search if submitting for a peer-reviewed version of the article?

On 2018-06-14 23:28:31, user Jeff Ellis wrote:

This reminds me of the problem of distinguishing host species specific pathovars of Fusarium oxysporum with DNA based diagnostics. Years and dollars were spent tweaking PCR assays directed at house keeping genes and ribosomal RNA spacers to no avail. In Australia the answer to Fov diagnosis for cotton and more recently Tropical Race 4 of banana has been to exploit the large differences in effector gene sequences between pathovars to develop truly diagnostic markers. Jeff Ellis

On 2018-05-15 07:05:45, user Hannah Gruner wrote:

Fascinating work!

I’m intrigued what signaling pathway Ddx3x is important for given the lack of change in canonical Wnt signaling. I’d be curious if non-canonical Wnt signaling may be involved considering the PCP pathways role in cortical neuron maturation (PMID:26939553; 19332887).

The intermediate progenitor increase in the Ddx3x LOF reminds me of a similar phenotype observed Slit1/2 and Robo1/2 mutant animals (PMID:23083737). As Slit-Robo signaling is associated with intellectual disabilities (PMID:12195014), and Slit2 and Robo1 are highly enriched in the Oh, et al. 2015 iCLIP data, it would be interesting to see if this signaling pathway is affected in Ddx3x knockdown as well.

On 2018-02-16 12:46:21, user Helder Maiato wrote:

The study by my esteemed colleague Dr. David Pellman and colleagues on the mechanism of nuclear envelope defects on lagging chromosomes has been kindly shared with me by the author, with whom we have been comparing results and discussing data interpretation. For the sake of transparency, I am convinced that sharing our points of convergence and divergence with the community is the correct way to move forward and widen the discussion. In summary, the manuscript by Dr. Pellman and colleagues confirm that lagging chromosomes in anaphase are defective in the recruitment of “non-core” nuclear envelope (NE) proteins, including nuclear pore complexes (NPCs), as shown by others, including our lab (e.g. see Afonso et al., Science, 2014; PMID: 24925910). Interestingly, they now look at “core” NE proteins and found that they are normally recruited to lagging chromosomes. As result of defective NE assembly, they also confirm that micronuclei derived from lagging chromosomes have impaired nuclear import and fail to accumulate important nuclear proteins required for genome integrity (e.g. RPA). A key question is what causes NE assembly defects on lagging chromosomes. The model proposed by Dr. Pellman and colleagues favors that spindle midzone microtubules, namely their bundling, inhibits non-core NE assembly, in an Aurora B-independent manner. This is precisely the point of debate, because we have previously shown that Aurora B inhibits NE assembly, depending on chromosome positioning along the spindle axis during anaphase, in both Drosophila and human cells in culture (Afonso et al., Science, 2014; PMID: 24925910). <br /> 1) Main points of convergence:<br /> - we confirm the spatial differences between core and non-core NE proteins on lagging chromosomes in human cells. This is a chapter of my former student Olga Afonso PhD thesis that is publicly available (a copy can be requested at: https://repositorio-aberto....<br /> - we also agree that early inhibition of Aurora B activity at anaphase onset compromises spatial regulation of NE assembly on lagging chromosomes in human cells. However, Aurora B inhibition at later time points after anaphase onset does not impair NE assembly on lagging chromosomes. Our interpretation is that this treatment, meant to be acute, might not be sufficiently fast to revert the process if performed too late in anaphase.<br /> - SMC2 association with chromosomes is also spatially controlled in human cells (Dr. Pellman’s data), i.e. lagging chromosomes show higher Condensin association, as we reported for S2 cells. Note that WE HAVE NOT LOOKED AT CONDENSIN LOCALIZATION IN HUMAN CELLS in our original study.<br /> - both studies agree that completion of NE assembly is not a global event, as it would be predicted by a “clock” model, supporting local regulation at the individual chromosome level.

2) Main points of divergence:<br /> - Loss of spatial regulation of NE assembly on lagging chromosomes after MKLP2/Subito RNAi. In S2 cells, this is lost in 90% of the cases we have looked at directly by live-cell imaging of Nup107-mRFP with 30 sec resolution. For us, this was the killer experiment for a role of Aurora B at the midzone in this spatial control of NE assembly. WE HAVE NOT DONE THIS EXPERIMENT IN HUMAN CELLS in our original study and are currently assessing the effect of MKLP2 inhibition in human cells. Thus, system-specific differences cannot be excluded.<br /> - In S2 cells, after RNAi of the non-SMC CAP-H/Barren/Condensin I subunit, spatial control of NE assembly is lost, as in Aurora B inhibition at anaphase onset. The work of Dr. Pellman now reports that SMC2 is not required for spatial control of NE assembly in human cells. We wondered whether redundancy between human Condensin complexes/subunits could explain the reported differences?<br /> - The model proposed by Dr. Pellman and colleagues does not explain the position dependent gradient of phosphorylated Histone H3 on S10 [PH3(S10)] on anaphase chromosomes (including laggards).

3) Specific comments on the manuscript by Dr. Pellman and colleagues:<br /> - Page 1, lines 28 and 29: “Non-core” proteins assemble at the spindle periphery (Extended Daata Fig. 1a) – would be helpful to provide microtubule staining to infer spindle position.<br /> - Page 2, lines 64-66: the role of an Aurora B phosphorylation gradient in the incorporation of lagging chromosomes into the main chromosome mass, as proposed in our paper by Afonso et al, is a corollary of the proposed function of the gradient in the spatial regulation of NE assembly during normal anaphase, but it is not exempt from failure, as discussed thoroughly by Maiato et al., Bioessays, 2015; PMID: 25470791. In fact, we report that, for S2 cells, only 50% of lagging chromosomes are able to integrate the main nuclei. Thus, the Aurora B gradient is a facilitator, not a guarantee (as for any other checkpoint).<br /> - Page 3, lines 70-71: Chromosome bridges are uniformly depleted for non-core proteins, arguing against a role for the Aurora B phosphorylation gradient. However, as we showed in our paper by Afonso et al., the Aurora B phosphorylation gradient is not smooth, and rather undergoes a sharp transition from mid to late anaphase (see Fig. S4B’ in our paper). This suggests that complete NE assembly is determined by a phosphorylation threshold, rather than a stepwise transition.<br /> - Page 3, lines 77 and 78: main prediction of the checkpoint model as a monitor of the position of lagging chromosomes. We never claimed that a chromosome separation monitors or detects lagging chromosomes (see point above). What we claimed is that a chromosome separation checkpoint monitors normal chromosome separation during anaphase. The lagging chromosome situation might benefit from this mechanism, but again, is not exempt from failure.<br /> - Page 3, lines 81-88: partial synchrony and Aurora B inhibition at all intervals from metaphase through telophase. This is an interesting experiment, which unfortunately does not allow precise control of Aurora B inhibition. The fact that no effect was observed when Aurora B inhibitor was added at later time points, might simply reflect that there was not enough time for the cellular uptake of the inhibitor to be effective.<br /> - Page 4, lines 116-120: effect of nocodazole treatment similar to Aurora B inhibition. This shows that the establishment of the Aurora B phosphorylation gradient is dependent on microtubules. <br /> - Page 4, lines 127-136: Taxol treatment upon Aurora B inhibition was used as the key experiment to exclude a role of Aurora B in the assembly of “non-core” proteins. However, it should be noted that both drugs were added 4 min after anaphase onset, and thus Aurora B inhibition might not have been effective, as shown previously by the authors. Several critical controls are missing in this experiment: a) microtubules are not labeled, which makes it difficult to evaluate the effect of both drugs on spindle architecture and positioning; b) where is Aurora B after the drug treatments?; and finally c) Aurora B activity on the lagging chromosomes should have been monitored [e.g. PH3(S10)]. WE HAVE NEVER SAW A PH3+ CHROMOSOME THAT RECRUITED NON-CORE ENVELOPE PROTEINS, INCLUDING NPCs, IN ALL CELL TYPES WE HAVE LOOKED AT (Drosophila, human, Pig, Muntjac, PtK), suggesting that Aurora B inhibits proper NE assembly.<br /> - Page 5, lines 140-142: KIF4A knockdown preserves spindle organization and central spindle localization of Aurora B (Extended Data Fig. 7a). The example provided actually shows remarkable differences in Aurora B localization in the central spindle (e.g. Aurora B is clearly more dispersed, as it would be expected due to less MT bundling after KIF4A RNAi). Also in Fig. 3b Aurora B localization should have been monitored because central spindle MTs look less dense after KIF4A RNAi.<br /> - Extended Data Figure 4b: This is another critical point of debate, hence important to be extra careful. In the example shown after MKLP2 RNAi the localization of Aurora B is surprising, because it should have remained extremely bright on anaphase chromosomes, but it is generally dispersed. Would be critical to monitor Aurora B activity [e.g. PH3(S10)] on lagging chromosomes after MKLP2 RNAi.<br /> - Extended Data Figure 5b: in the bottom panel, it looks that the contrast has been adjusted differently between the low mag panels 1-3) and the corresponding high mag panels (particularly panel 2). Maybe worth double checking for a fair comparison.<br /> - Extended Data Figure 8: I don’t really understand what is going on here. Maybe worth including a picture of the whole cell and a bit of the history of that cell in life before forming micronuclei?<br /> - more general comments: I didn’t quite see the evidence that links NE defects on lagging chromosomes with chromothrypsis in this manuscript, but I might have missed something. It is also unclear how microtubules would selectively prevent the assembly of non-core NE proteins on lagging chromosomes. Given that microtubules control Aurora B activity and will likely be critical to establish the dimensions of the Aurora B phosphorylation gradient [e.g. see Noujaim M, Bechstedt S, Wieczorek M, Brouhard GJ (2014) Microtubules accelerate the kinase activity of Aurora-B by a reduction in dimensionality. PLoS One 9:e86786. doi:10.1371/journal.pone.0086786], it looks to me that it is extremely difficult, if not impossible, to separate both variables, i.e. microtubules and Aurora B (reminds me of the long-lasting debate between tension vs. attachment for the spindle assembly checkpoint). Overall, we stand by every single piece of data that we published in the Afonso et al. paper, which, in our opinion, does not exclude that MTs might inhibit NE assembly (e.g. by steric hindrance). In my view, it will be important to account for possible differences between systems and the possible communion, rather than exclusion, of both models. <br /> This is my first post in BioRxiv, which I believe is meant to help authors to improve their manuscripts and receive broad feedback prior to publication, while fostering the discussion with a larger community. I hope the authors find our comments useful and that other people can join the discussion, and, worst comes to worst, we agree to disagree on some points!

With best regards and good luck with your paper,<br /> Helder Maiato

This section on patriarchy stood out to be me cause it explains how the family structure was historically used by men to control women as economic and sexual property. It’s crazy to me that only a century ago, a wife’s earnings belonged to her husband and not to her. Even today, women still handle the majority of housework and childcare. This reminds me of the social dynamics I've seen in my favorite shows.

This section on patriarchy stood out to me because it explains how the family structure was historically used by men to control women’s as economic as sexual; proper. It’s crazy to think that only a century ago, a wife’s paycheck belonged to her husband. Even today, women still handle the majority of housework and childcare. This reminds me of some of my favorite shows that I like to watch at home.

This line from the reading stood out to me because of how private messaging on social media isn’t actually fully private and feels like an invasion of privacy. It makes me think about how often we trust platforms with personal conversations without really considering who else might have access. From a user perspective, there’s a tradeoff in how we want privacy, but we also want safety features like being able to report harassment or threats. That tension reminds me of discussions in this course about data mining and surveillance, where companies justify access to data for protection or improvement, but it can also blur ethical boundaries. I also wonder where the line should be drawn for these social media companies ethically.

This is interesting because it reminds me of what I did in my art appreciation class during my sophomore year at crowder college. Giving the students an image and having them interpret their individual feelings and understandings related to the image as part of a group discussion is a great exercise. As the old saying goes "a picture is worth a thousand words" and this is especially true when it comes to looking at art. And if its an art that an ML student could understand from their cultural heritage then they can share that understanding and unique interpretation with their non-ML classmates in their group.

These statements highlight how confidence and identity play a major role in learning, especially for multilingual learners. This reminds me of growth mindset because it focuses on effort rather than ability, and I think teachers can build this by creating inclusive environment where all students feel valued.

This selection reminds me of the times when segregation in schools was slowly changed into integration and many students of different ethnic and cultural background started joining white americans in schools. Because there was sadly a lot of controversy and fear back that that the african american, hispanic, muslim, etc. would affect the school's academics, decrease home values in white neighborhoods, and take away jobs. Even still to this day there are still some community members that are so set in that older systemic social construct because it was all they knew growing up. And it was because they never took the time to get to know those families and their culture.

This part of the chapter really stood out to me because if you look at enough data, you can basically find support for almost any conclusion or interpretation you could want. This made me think about how often I see graphs and statistics online and just assume they mean something important without really questioning them. It reminds me of what we've been learning about data visualization and how easy it is to make something look convincing depending on what data you choose to show. It also made me realize that data minding isn't just technical but also ethical as well. If people can findpatterns that match what they already believe, then it's easy to spread misleading ideas without realizing it. I wonder if platforms should do more to limit this kind of thing or if its just on users to think more about the data they see online.

I knew after reading the "Your Friends’ Online Connections Can Reveal Your Sexual Orientation" article that researchers use shadow profiles to predict, if not saying exposing, personal information of non-users. This actually scares me rather than surprises me as it shows that users' privacy can be indirectly exploited just from their contact lists. Even when it's not guaranteed that this information is completely accurate, it concerns both social media users and non-users as it's now known that their personal information can be "inferred" and potentially sold for marketing firms. This reminds me of the Facebook's 5-billion penalty regarding their violation of users' privacy that leads to Facebook changing their personal information protection terms and conditions.

This is a very interesting idea to me because If a memory is being used as a truth, how true is it really? It reminds me of playing a game of telephone in which your memory can warp with time. I am guilty of misremembering things too so writing out my thoughts can be difficult if its not fresh in my head.

This was suprising too me, but not something I would have expected to happen. I guess I am most surprised that social media or internet sites don't censor this more. This reminds me of the Westboro Baptist Church and how they would mock families at military funerals.

Light on what he did, but interesting composition. Reminds me of [[Mijn intellectuele stamboom]]

I love this statement because it reminds me of a spiral curriculum, one where learning doesn’t stop until we know students have truly mastered it. It requires us to continuously examine evidence and data, monitor progress, and adjust our lessons to ensure students can transfer their learning and engage in deep, meaningful understanding.

This reminds me of the "Do No Harm" ethics of medicine.

I have not seen this vocab but the idea makes sense. Reminds me of my class last summer about transparency with populations being studies to include them on the research itself too.

Very good points here. Reminds me of similar points from Kate's OSR blog. The one about NPCs being independent. The difficulty I would say s the cognitive load of it all. I think most people want an interesting and engaging NPC but keeping up with that can be quite draining and is a skill I dont think everyone necessarily has. They might be able to do it but it's not easy for them.

This reminds me of one of the conversations we had as a class, on the idea that once a society has been introduced to capitalism, there is essentially no way of going back. I wonder how you think a utopia of sufficiency could illicit such a huge cognitive shift. As it stands today, it feels almost impossible to imagine the ecological take precedence over comfort. I wish for this utopia to be a reality, even if not wholly, having aspects of it would be wonderful.

Great parallel to B + F's Shock of the Anthropocene. It reminds me of our class discussion on the power of the "shock factor" within Anthropocene discourse.

I see the motivation for why they believe that white must take over the world. They think that this is a race war. That only one race will conquer the world and subjugate everyone in the process and White People MUST take up arms together against this threat. 

It reminds me a lot about the Russia and American space race. Where each group were competing with each other, even though they should have been working together... pretty petty if you ask me. 

Author response:

Reviewer 1:

Porte et al. investigate how observers form confidence judgments about the presence vs absence of near-threshold audiovisual stimuli. In two psychophysical detection experiments, human participants judged whether a stimulus (visual, auditory, or audiovisual) was present or absent, reported amodal confidence, and then gave modality-specific detection and confidence ratings using a bidimensional scale. The authors report that audiovisual (AV) stimuli are detected more accurately than unimodal stimuli, but that multisensory stimulation does not improve metacognitive efficiency. Participants are more confident in absence than in presence judgments. They extend a previously proposed model to an audiovisual setting, assuming evidence is available only for presence and that absence is inferred via counterfactual detectability. Detection is modeled with a disjunctive integration rule across modalities, while confidence is explained by a combination of conjunctive (for presence) and disjunctive/negation-of-disjunction (for absence) rules.

We thank the reviewer for thoroughly evaluating our work.

There are several points I wish to have clarified, outlined below:

(1) Framing of bimodal vs unimodal detection

On p.3, the introduction states that "Adults typically show higher detection rates and faster reaction times for bimodal than for unimodal stimuli." This is broadly consistent with the literature, but as written, it obscures the fact that these effects depend critically on experimenter-defined stimulus strengths. It is trivial to construct cases where a strong unimodal stimulus is more detectable than a bimodal stimulus made of two very weak unimodal stimuli. If "bimodal" is understood as the co-presentation of two unimodal components matched in detectability, then Bayes-rule-based arguments indeed predict better detection for the bimodal case; how much better is theoretically interesting, but not quantified in this paper. There is an entire literature on the combination of two unimodal stimuli, which is not touched on. For a pertinent reference, see Ernst & Banks 2002. I recommend clarifying that the statement assumes comparable unimodal intensities.

We will clarify that when discussing bimodal stimuli, we mean the co-presentation of two unimodal stimuli of similar intensity. We will add references to the literature during discrimination tasks that have shown that multisensory cue-combination followed Bayes rule integration (e.g., Ernst & Banks, 2002; Battaglia et al., 2003; Alais & Burr, 2004) and clarify in which ways our work differs from this rich body of work and provides novel contributions.

(2) Relationship to signal detection theory and counterfactual perceptibility

In the introduction, the authors write, "If sensory evidence is only available for presence," motivating counterfactual perceptibility as a necessary ingredient to infer absence. However, standard signal detection theory (SDT) already provides a widely accepted framework in which a continuous internal response is present on both signal and noise (absent) trials, with absence corresponding to the noise distribution and decisions implemented by a criterion. Thus, there is no logical need to invoke counterfactual perceptibility simply to define absence; rather, the Mazor-style framework adds an explicit belief model about detectability and an optimal stopping policy. It would strengthen the paper to more clearly state how the proposed model goes beyond SDT conceptually, acknowledge that SDT can account for presence/absence decisions without counterfactuals, and position the counterfactual account as a hypothesis about how observers actually compute absence/confidence, not as a necessity.

One of the central claims of the paper is that detection in the case of absence requires counterfactual reasoning. The authors should demonstrate whether or not an SDT-based generative model can describe these amodal and uni- and bi-modal stimulus decisions. In such an SDT model, an SDT-based generative model in which the noise distribution is shared across conditions, and unimodal vs bimodal differences are captured by changes in the mean or variance of the signal+noise distribution.

We will clarify that our framework explains how absence judgments (and related confidence) are formed, and what it adds to SDT models, including the reproduction of reaction times and a normative explanation of criterion placement (results about RTs are available in the supplementary materials).We will also run additional model comparisons assessing how an SDT-based generative model performs compared to our Bayesian model based on counterfactual perceivability.

(3) Confidence vs performance: is AV confidence special?

The paper's central claims about multisensory confidence and metacognition would be stronger if the authors showed that AV confidence deviates from what is expected given performance alone. From the reported results, AV accuracy is around 80%, with visual and auditory at about 60% and 40%, respectively. Given that confidence typically monotonically scales with accuracy, the first question is whether AV confidence is entirely explained by improved performance, or whether there is an additional multisensory contribution. A simple, informative analysis would be for each subject, plot mean confidence vs per cent correct for AV, V, A, and absent conditions, and to test whether AV confidence lies above the trend predicted by accuracy alone.

This is an excellent suggestion, and we will conduct the proposed analysis.

(4) Metacognitive measures: logistic regression slopes vs meta-d′/d′

In the "Multisensory effects on metacognitive performance" section, the authors define "metacognitive sensitivity" as the slope of a Bayesian logistic regression predicting accuracy from confidence. There is substantial literature showing that logistic-slope measures of metacognitive sensitivity are criterion-dependent and can be affected by both task and confidence criteria (for one example, see Rausch & Zehetleitner, 2017). In contrast, meta-d′/d′ was specifically developed to provide a bias-invariant measure of metacognitive efficiency. Though this, too, is dated (see Boundy-Singer et al., 2023). Given that the authors already estimate HMeta-d-based M-ratios, it is unclear why they rely on logistic regression slopes as their primary "metacognitive sensitivity" metric in Figure 4A. I suggest either replacing the logistic-slope metric with SDT-based measures (meta-d′, meta-d′/d′) or providing a clear justification for using logistic slopes, along with a discussion of their known limitations.

Additionally, Figure 3 reports M-ratios without showing the corresponding d′ or meta-d′ for judge-present vs judge-absent conditions. Presenting these would help contextualize the metacognitive efficiency results and clarify whether differences are driven mainly by changes in metacognitive sensitivity, changes in task performance, or both. The d' values per condition could be added to Figure 2A.

All typical measures of metacognitive sensitivity are influenced by metacognitive bias and task performance to some extent, and none of them is a pure measure of type-2 sensitivity (e.g., see Rahnev, 2025). Here, we chose logistic regression because it enables modeling interactions with other predictors in a factorial design with a limited number of trials.

We will clarify the limitations of metacognitive sensitivity measures and better explain why we then used Mratio to estimate metacognitive performance while controlling for underlying task performance.

Thank you for this suggestion. We will add the d’ values per condition to Figure 2A.

(5) Interpretation of confidence in absence vs presence

The authors emphasise that it is surprising subjects are more confident in absence than in presence judgments, both at amodal and modality-specific levels. However, Figure 2B suggests that absent responses are very accurate: absent is reported as present only in about 10% of absent trials, implying a high correct rejection rate. If confidence tracks outcome probability, higher confidence for absence may be at least partly expected. Before attributing this asymmetry primarily to counterfactual reasoning, it would be important to explicitly relate confidence to accuracy for hits, misses, false alarms, and correct rejections and show whether absence confidence remains elevated relative to presence after controlling for accuracy differences across judgment types and conditions. Without this, the interpretation that higher absence confidence is inherently "unexpected" seems overstated.

This higher confidence for absence judgments than for presence judgments was observed while controlling for response accuracy. We will clarify this in the main text.

(6) Model: integration rules, confidence, and evidence strength

The modeling section extends the Mazor et al. ideal observer to two modality-specific sensors, with disjunctive integration for detection and then disjunctive vs conjunctive integration rules for confidence. I have a few comments.

First, the detection rule is disjunctive and is reported as a finding. However, the conclusion that detection relies on a disjunctive rule ("present if A or V") closely mirrors the task instructions-participants are explicitly told to respond "present" if they detect the stimulus in any modality. As such, this seems more like a sanity check than a novel empirical finding. Relatedly, the conjunctive detection is a weak null. The conjunctive rule ("present only if both A and V") is behaviorally implausible given the task instructions. A more informative baseline would be an SDT-style scalar-evidence model (see comment 2), rather than a conjunctive rule that participants would have to actively violate the instructions to follow.

Second, confidence in the model is defined as the probability of being correct at the time of the detection decision. However, this implies a fixed amount of evidence at decision time unless additional mechanisms are invoked. This issue is well known in diffusion modeling (see Kiani et al. 2014) and deserves explicit discussion; otherwise, it is unclear how the model produces graded confidence from a bound-crossing rule alone.

Third, the authors do not consider a straightforward evidence-strength account of confidence. When both modalities indicate presence, there is, on average, more total sensory evidence than in unimodal trials, making correct decisions more likely and, under most frameworks, confidence higher. Likewise, weak evidence in both modalities can be stronger evidence for absence than moderate in one and weak in the other. Many of the patterns that motivate the presence-conjunctive/absence-disjunctive mix could arise from a model where confidence simply reflects the amount of evidence for the chosen option, without positing distinct logical integration rules for presence vs absence. As the authors note, purely disjunctive or purely conjunctive confidence rules fail to capture the trends in confidence reports in Figure 7, leading them to adopt a combined presence-conjunctive/absence-disjunctive rule. A more parsimonious alternative-that confidence scales with evidence magnitude and cross-modal agreement-should be explicitly considered and, ideally, implemented as a competing model. Finally, if the model is intended as a good account of the data, it would be useful to report whether it also reproduces the metacognitive efficiency patterns (M-ratios) beyond the mean confidence patterns shown in Figures 7-8. At present, the model appears systematically over-confident, which should be acknowledged and quantified.

Indeed, the disjunctive rule was expected, given our design; we will clarify this. As mentioned above, we will directly compare the results of our current model with those of a more traditional SDT-based generative model, as suggested by the reviewer.

Contrary to a classical drift diffusion model, the model does not assume a fixed decision boundary, but derives an optimal stopping policy per time point and belief state. As a result, and depending on beliefs about perceptual evidence and the temporal discounting factor, optimal decision boundaries can be asymmetric and may collapse asymmetrically toward 0. Furthermore, given the asymmetry in the information value between sensor activations and inactivations, and differences in the information value of sensor activations of the two modalities, boundary crossing can lead to belief states that are far or close to the decision boundary, depending on the nature of the evidence. Together, even without an explicit modeling of post-decisional evidence, the model can account for variability in the total accumulated evidence at decision time.

From our understanding, the proposed alternative is equivalent to our current model, in which confidence scales with evidence magnitude.

The model was not fitted to confidence data, which could explain its overall overconfidence. To further test our model, we will assess its ability to reproduce patterns of metacognitive efficiency (M-ratios).

(7) Confidence asymmetry index (CAI) and modality weighting

The confidence asymmetry index (CAI) is defined as the difference between auditory and visual confidence on AV vs absent trials, and the authors report strong correlations between observed and simulated CAI across participants. They interpret this as evidence that subjects place different weights on auditory vs visual signals. Several questions arise. First, does CAI capture asymmetries beyond what is expected from accuracy differences between modalities and conditions? Second, because the simulated data are generated from model fits to the observed data, a correlation between observed and simulated CAI is expected: the model is built to reproduce the individual patterns it is then compared to. A stronger test would compare CAI from data simulated with modality-specific belief parameters, versus CAI from data simulated with constrained equal belief parameters (same θs). Relatedly, the paper would benefit from a plot showing the distribution of θs for A and V- present stimuli across subjects. These values could also be related to unimodal sensitivity measured in the calibration/training phases. A natural prediction is that higher unimodal sensitivity should correspond to higher belief parameters for presence.

The model was not fitted to either the modality-specific responses or the confidence ratings, so the correlation between observed and simulated CAI was not expected and provides a good test of our model's ability to reproduce the observed patterns. We will test whether the same correlations hold when using the difference in accuracy instead of the confidence.

We found that the best model is the one with the same belief across the visual and auditory sensors. Given this, we cannot investigate how modality-specific belief parameters are linked to unimodal sensitivity for each participant.

Reviewer 2:

Summary:

In this study, across two experiments, the authors wrestle with the question: What is the profile of confidence judgments in presence/absence decisions for audiovisual stimuli? After thresholding observers to 50% target detection rates in each modality, the authors conducted one experiment that included 75% target presence (spread equally across bimodal, auditory, and visual targets) and one experiment with 50% overall target presence. Results showed that, overall, detection performance was higher for audiovisual stimuli compared to unimodal ones, and that a recent model for stimulus detection could be extended to this multisensory scenario. By incorporating a disjunctive rule for absence judgments and a conjunctive rule for presence judgments, the model was able to qualitatively reproduce some of the trends observed in the human data regarding confidence.

Strengths:

(1) The paper makes novel contributions to the study of multisensory confidence judgments for yes/no target detection.

(2) The paper further extends the use of a leading model of stimulus detection (from Mazor et al., 2025).

(3) Pre-registration of the study was implemented, and the code is publicly available (although the GitLab link requires registration to access the materials).

(4) One of the empirical results (higher confidence for absence compared to presence judgments) is especially interesting, contributing another empirical finding to a very mixed literature on this topic (as the authors note).

We thank the reviewer for the positive evaluation of our work.

Weaknesses:

(1) Page 5 - I have concerns about the use of the equal-variance model from Signal Detection Theory to analyze the data. For example, the authors should read the recent paper by Miyoshi, Rahnev, and Lau in iScience, found at this link: https://www.cell.com/iscience/fulltext/S2589-0042(26)00373-1 . In this paper, the authors note how the equal variance model should be used with caution in yes/no detection tasks, since the variances of the "stimulus present" and "stimulus absent" distributions are often different from one another. In a revision, I highly recommend that the authors explicitly discuss this paper and review whether the assumptions for the equal-variance model have been met (e.g., since they have confidence data, one way to do this would be to evaluate if the slope of the line in zROC space differs from 1). The authors may also want to incorporate methods from this iScience paper into the current manuscript, or potentially move to using an unequal variance SDT model and compute d'a and c'a.

This is an excellent suggestion. We will run this analysis and refit the d’ and criterion response using unequal-variance models to see whether we observe the same results.

(2) Related to the computation/measurement of the response criterion, the authors note on page 18 in the Methods that for Experiment 1, signals are actually present on 75% of trials, since a bimodal stimulus is present on 25% of trials, the visual circle only occurs on 25% of trials, the sinusoidal tone occurs on 25% of trials, and then only noise is present on 25% of trials. Did the authors have any a priori hypotheses about the response criteria that participants would exhibit in Experiment 1, considering the unbalanced target presentation rate in this task? Also, in Experiment 2, what did it mean to equate target present and target absent trials? Is it that they broke 50% target present trials down into 16.67% bimodal targets, 16.67% visual targets, and 16.67% auditory targets? A few more details would be good to explicitly note for those trying to replicate the task

We will clarify this point in the manuscript. In Experiment 2, the stimulus was absent on 50% of the trials. As a result, the 50% of stimulus present trials were split into the three possible conditions, resulting in a sixth of the trials being auditory, a sixth visual, and a sixth audiovisual; we will make these proportions clearer in the text.

We did not have any a priori hypotheses about the response criteria for Experiment 1. The reviewer is right, the proportion of absent versus present trials can indeed have an impact on response bias. In fact, one of the goals of Experiment 2 was to test whether the low frequency of absent trials compared to present ones could explain both response bias and higher confidence in absence observed in Experiment 1, which we found was not the case, as we did not observe a difference between the two experiments. We will clarify this in our revision.

(3) It is important to plot the individual data for Figure 2. If the authors didn't match detection performance for the visual and auditory modalities, it would be good to see the individual data to know why. Is it that the thresholding procedure didn't work for some of the participants in the visual modality, and that's why the "yes" response rate is (on average) ~60% or higher across the two experiments? Similarly, in the auditory domain, do the authors have participants that are at floor? Or is it simply that the staircases failed to successfully target 50% detection on average?

We will add individual data to Figure 2.

Indeed, staircases failed to achieve 50% detection on average; participants for whom psychometric curves did not converge were excluded, as were those at floor level in one of the two modalities.

(4) The authors mentioned that data were collected on the Prolific platform. What checks did they conduct to ensure that this data wasn't produced by bots? There are recent high-profile publications in PNAS and Behavioral Research Methods that indicate how online data collection is problematic (e.g., https://www.pnas.org/doi/10.1073/pnas.2535585123and https://link.springer.com/article/10.3758/s13428-025-02852-7 ). What analyses or quality checks are there to ensure that humans were the ones completing the task?

Data were collected on the Prolific platform, which has been shown to yield high-quality data (Kay, 2025). However, we agree that this is a potential concern and will add a note of caution in the revised manuscript, even if the risk that the data do not come from humans but from bots is low (Huskey et al., 2026; Chetverikov, 2026).

(5) Page 7 - Since confidence was collected on a continuous scale, the authors should say a bit more about how they were able to compute measures of metacognitive efficiency. My understanding is that to compute meta-d', the data has to be binned. How was the binning implemented? With whatever bin size the authors chose, would it make any difference to the results if they changed the number of the bins in the analysis?

We will clarify this aspect of the analysis. Data were binned into four quartiles based on the overall distribution of confidence values across participants, based on the binning used in the example in Fleming (2017). We will examine whether changing the number of bins changes the results (Dayan, 2023).

(6) Page 8 - Is there a prior precedent for using slope of the Bayesian logistic regression predicting accuracy from confidence as a measure of metacognitive sensitivity? If so, can the authors cite those papers as a reference? If not, can they place this analysis within the context of other measures of metacognitive sensitivity that exist? (meta-d', AUROC (Type 2), etc.)

Yes, logistic regression has been used to quantify metacognitive sensitivity before. We will add the relevant papers as references (e.g., Sandberg et al., 2010; Norman et al., 2011; Siedlecka et al., 2016; Wierzchoń et al., 2012; Faivre et al., 2018; Pereira et al., 2023)

(7) Page 8 - Another one of the results on page 8 is worth reflecting further upon: the authors note how in Experiment 1, no credible difference was found between unimodal and bimodal trials (DeltaM = -0.25 [-0.59, 0.10]), but in Experiment 2, "we observed higher metacognitive efficiency in unimodal compared to bimodal trials (DeltaM = -0.28 [-0.54, -0.02]. Those DeltaM values are nearly identical, so without a power analysis motivating the number of participants the authors collected, how certain are they that the results from these two experiments are really that distinct? It reminds me a bit of the Andrew Gelman blog post, "The difference between significance and non-significance is not significant".

The number of participants was determined using a Bayesian optional stopping rule, as preregistered. The reviewer is right that the delta values are very similar in the two experiments. Given that a difference was found in only one experiment, we decided not to draw conclusions from it.

(8) Is there any way to look at whether the presence of multisensory hallucinations (or perhaps that word is too strong, and we should simply consider them miscategorizations) increased as the task progressed? That is, the authors have repeated presentations of audiovisual stimuli for at least some percentage of the trials. Since the percentages for auditory stimuli being correctly categorized as auditory are at 85% in Experiment 1 and 79% in Experiment 2, were the trials where they miscategorized these stimuli equally spread throughout the task? Or did they come later in the experiment, after being repeatedly exposed to multisensory trials?

We will examine how the proportion of miscategorisation changed throughout the task.

(9) Would the authors obtain the same results if they got rid of the amodal confidence judgment in their task, and simply had participants report the bimodal confidence following the presence/absence judgment? Part of the reason for asking this is that, according to page 11, the model is only fitted to amodal detection accuracy and response time data. This surprised me. I would have expected that the bimodal confidence would provide more useful information for the model fit. The authors should further explain this rationale in the paper. It seems odd to me to have the multisensory confidence ratings and not have them play a central role in the modeling work.

Our main goal was to investigate how participants form integrated, supramodal confidence judgments on the basis of multisensory sources of information. Therefore, the amodal confidence judgments are required here.

Moreover, the model was fitted to response times that corresponded to the amodal judgment. Because we had no meaningful response times for the modality-specific judgment, we could not use them to fit the model.

(10) In Figure 6, it appears the model is a bit off in its estimate of auditory responses (panel B, E) in the AV condition. Do the authors have any intuitions about why this might be happening?

Indeed, the model does not capture the full behavioral effects reflecting multisensory interference in the modality-specific responses. We suppose that the model does not reproduce these interferences, as it is only fitted to amodal detection accuracy, and as the two sensors are completely independent from one another. We will clarify this aspect in the text.

(11) The authors talk about how the model is reproducing effects in the human data, but there's no systematic comparison, quantitatively, of how the two things relate. The authors should include some quantitative measure that reflects this

In addition to the d’ and criterion comparison between the observed and simulated data, we will compare modality-specific d’ and the correlations between observed and simulated confidence.

(12) Related to this, I am not sure I agree with the characterization in Figure 7 that "when confidence followed a disjunctive rule, the model failed to capture important aspects of the data. On the other hand, when confidence followed a conjunctive rule, it reproduced confidence in presence judgments but failed to capture variability in confidence ratings for absence judgments." What, quantitatively, is the basis of this claim? This applies to Figure 8, too. I am not clear how, specifically, and quantitatively, the authors are justifying their claims about model fits. I don't think the confidence asymmetry index in Figure 8 is enough to quantify the quality of the model fitting procedure.

To further support this claim, we will add a quantitative comparison of the different confidence fits.

(13) Is there any chance the higher metacognitive efficiency for auditory trials is simply driven by differences in the d' values across the modalities? It might be good to probe this effect further.

Thank you for this remark. Indeed, the difference in metacognitive efficiency may be driven by differences in the d’ values, and so a lower d’ for auditory stimuli can lead to higher metacognitive efficiency for a similar metacognitive sensitivity.

Reviewer 3:

This study used a pre-registered novel behavioural paradigm and computational modelling to investigate multi-sensory influences on detection and confidence. Participants performed amodal detection of auditory and visual stimuli (indicating that a stimulus was there when either an auditory stimulus or a visual stimulus or both were present), followed by amodal and unimodal confidence ratings. Detection was higher when both stimuli were present, and the presence of one modality increased the confidence in the presence of the other modality. In contrast to previous detection studies, confidence was higher for absent than for present judgements, but metacognitive efficiency was higher for present judgements. Metacognitive sensitivity was higher for bimodal stimuli, but this was not the case for metacognitive efficiency, suggesting that the sensitivity might be driven by first-order performance. The computational model showed that both detection and confidence in absence followed a disjunctive evidence integration rule, while confidence in presence followed a conjunctive integration rule.

We thank the reviewer for engaging with our work.

Strengths:

The paper has several major strengths. Firstly, it addresses a novel research question using an innovative and well-controlled paradigm. Furthermore, the paradigm and analyses were pre-registered, and all effects that were interpreted were replicated in two independent samples. Finally, the paper uses an advanced computational model to capture counterintuitive patterns in the data.

Weaknesses:

The major weakness of the paper is the narrative structure. It is not always clear how the different analyses relate to the main research question. Many different effects are reported in terms of detection accuracy, bias, confidence and metacognition, as well as cross-modal and unimodal versus bimodal effects. It would help readability if the paper were streamlined in terms of the research question that is being answered, which I believe is specifically about multimodal absence judgements. Relatedly, for a reader not intimately familiar with the metacognition literature, the difference between MRatio, metacognitive sensitivity and metacognitive efficiency is not obvious. It would be good to clarify this more in the manuscript.

We will improve the narrative structure so that each result clearly relates to the research question.

We will also add a clearer definition of the various metacognition metrics to improve readability.

In general, the conclusions drawn by the authors seem to be supported by the results. However, I was missing quantitative model comparisons between the conjunctive and the disjunctive models and an explanation of why the models systematically overestimated the confidence ratings. Furthermore, the 'perceptual multisensory interference' section reports on very interesting effects, but these are not supported by statistical tests in the main text. It would help to assess the strength of the claims if the statistical evidence in favour of these claims were presented together in the main text.

The model was not fitted to confidence data, which could explain its overall overconfidence. As stated in previous responses, we will perform additional analyses to evaluate the model’s ability to reproduce confidence ratings. As some of the results were not replicated across experiments, we decided to put all statistical results related to multisensory interference in the supplementary materials and to focus only on consistent results across experiments.

One other concern is that in real-world multi-sensory perception, such as the mosquito example in the introduction, the auditory and visual signals have a strong natural association, which means that if you hear the auditory signal, you expect that you will see the visual signal soon and vice versa. As far as I understood, this association was not present in the current paradigm, which might influence the type of effects that one would expect to see.

The relation here is indeed artificial; we try to reinforce it as much as possible in the instructions of the task by indicating to the participants that they have to “detect a mosquito” that could be present auditory, visually, or both. But we acknowledge that the association between the visual and auditory stimuli is artificial, which may indeed influence our results.

References

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Chetverikov, A. (2026). Online behavioral studies are safe for now : Unusual RTs do not imply bots (A reply to Van der Stigchel et al., 2026) (Gjw5u_v1). PsyArXiv. https://osf.io/preprints/psyarxiv/gjw5u_v1/

Dayan P. (2023). Metacognitive Information Theory. Open mind : discoveries in cognitive science, 7, 392–411. https://doi.org/10.1162/opmi_a_00091

Ernst, M. O., & Banks, M. S. (2002). Humans integrate visual and haptic information in a statistically optimal fashion. Nature, 415(6870), Article 6870. https://doi.org/10.1038/415429a

Faivre, N., Filevich, E., Solovey, G., Kühn, S., & Blanke, O. (2018). Behavioral, Modeling, and Electrophysiological Evidence for Supramodality in Human Metacognition. Journal of Neuroscience, 38(2), 263‑ 277. https://doi.org/10.1523/JNEUROSCI.0322-17.2017

Fleming, S. M. (2017). HMeta-d : Hierarchical Bayesian estimation of metacognitive efficiency from confidence ratings. Neuroscience of Consciousness, 2017(1),

Huskey, R., Zhao, Z., Parry, D. A., & Fisher, J. T. (2026). An AI agent can complete the Attention Network Test with human-like behavioral signatures : Implications for the bot-or-not debate (T2jru_v1). PsyArXiv. https://osf.io/preprints/psyarxiv/t2jru_v1/

Kay, C.S. Why you shouldn’t trust data collected on MTurk. Behav Res 57, 340 (2025). https://doi.org/10.3758/s13428-025-02852-7nix007. https://doi.org/10.1093/nc/nix007

Norman, E., Price, M. C., & Jones, E. (2011). Measuring strategic control in artificial grammar learning. Consciousness and Cognition, 20(4), 1920-1929. https://doi.org/10.1016/j.concog.2011.07.008

Pereira, M., Skiba, R., Cojan, Y., Vuilleumier, P., & Bègue, I. (2023). Preserved Metacognition for Undetected Visuomotor Deviations. Journal of Neuroscience, 43(35), 6176‑ 6184. https://doi.org/10.1523/JNEUROSCI.0133-23.2023

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Siedlecka, M., Paulewicz, B., & Wierzchoń, M. (2016). But I Was So Sure ! Metacognitive Judgments Are Less Accurate Given Prospectively than Retrospectively. Frontiers in Psychology, 0. https://doi.org/10.3389/fpsyg.2016.00218

Wierzchoń, M., Asanowicz, D., Paulewicz, B., & Cleeremans, A. (2012). Subjective measures of consciousness in artificial grammar learning task. Consciousness and cognition, 21(3), 1141-1153. https://doi.org/10.1016/j.concog.2012.05.012

Reviewer #2 (Public review):

Summary:

In this study, across two experiments, the authors wrestle with the question: What is the profile of confidence judgments in presence/absence decisions for audiovisual stimuli? After thresholding observers to 50% target detection rates in each modality, the authors conducted one experiment that included 75% target presence (spread equally across bimodal, auditory, and visual targets) and one experiment with 50% overall target presence. Results showed that, overall, detection performance was higher for audiovisual stimuli compared to unimodal ones, and that a recent model for stimulus detection could be extended to this multisensory scenario. By incorporating a disjunctive rule for absence judgments and a conjunctive rule for presence judgments, the model was able to qualitatively reproduce some of the trends observed in the human data regarding confidence.

Strengths:

(1) The paper makes novel contributions to the study of multisensory confidence judgments for yes/no target detection.

(2) The paper further extends the use of a leading model of stimulus detection (from Mazor et al., 2025).

(3) Pre-registration of the study was implemented, and the code is publicly available (although the GitLab link requires registration to access the materials).

(4) One of the empirical results (higher confidence for absence compared to presence judgments) is especially interesting, contributing another empirical finding to a very mixed literature on this topic (as the authors note).

Weaknesses:

(1) Page 5 - I have concerns about the use of the equal-variance model from Signal Detection Theory to analyze the data. For example, the authors should read the recent paper by Miyoshi, Rahnev, and Lau in iScience, found at this link: https://www.cell.com/iscience/fulltext/S2589-0042(26)00373-1. In this paper, the authors note how the equal variance model should be used with caution in yes/no detection tasks, since the variances of the "stimulus present" and "stimulus absent" distributions are often different from one another. In a revision, I highly recommend that the authors explicitly discuss this paper and review whether the assumptions for the equal-variance model have been met (e.g., since they have confidence data, one way to do this would be to evaluate if the slope of the line in zROC space differs from 1). The authors may also want to incorporate methods from this iScience paper into the current manuscript, or potentially move to using an unequal variance SDT model and compute d'a and c'a.

(2) Related to the computation/measurement of the response criterion, the authors note on page 18 in the Methods that for Experiment 1, signals are actually present on 75% of trials, since a bimodal stimulus is present on 25% of trials, the visual circle only occurs on 25% of trials, the sinusoidal tone occurs on 25% of trials, and then only noise is present on 25% of trials. Did the authors have any a priori hypotheses about the response criteria that participants would exhibit in Experiment 1, considering the unbalanced target presentation rate in this task? Also, in Experiment 2, what did it mean to equate target present and target absent trials? Is it that they broke 50% target present trials down into 16.67% bimodal targets, 16.67% visual targets, and 16.67% auditory targets? A few more details would be good to explicitly note for those trying to replicate the task.

(3) It is important to plot the individual data for Figure 2. If the authors didn't match detection performance for the visual and auditory modalities, it would be good to see the individual data to know why. Is it that the thresholding procedure didn't work for some of the participants in the visual modality, and that's why the "yes" response rate is (on average) ~60% or higher across the two experiments? Similarly, in the auditory domain, do the authors have participants that are at floor? Or is it simply that the staircases failed to successfully target 50% detection on average?

(4) The authors mentioned that data were collected on the Prolific platform. What checks did they conduct to ensure that this data wasn't produced by bots? There are recent high-profile publications in PNAS and Behavioral Research Methods that indicate how online data collection is problematic (e.g., https://www.pnas.org/doi/10.1073/pnas.2535585123 and https://link.springer.com/article/10.3758/s13428-025-02852-7). What analyses or quality checks are there to ensure that humans were the ones completing the task?

(5) Page 7 - Since confidence was collected on a continuous scale, the authors should say a bit more about how they were able to compute measures of metacognitive efficiency. My understanding is that to compute meta-d', the data has to be binned. How was the binning implemented? With whatever bin size the authors chose, would it make any difference to the results if they changed the number of the bins in the analysis?

(6) Page 8 - Is there a prior precedent for using slope of the Bayesian logistic regression predicting accuracy from confidence as a measure of metacognitive sensitivity? If so, can the authors cite those papers as a reference? If not, can they place this analysis within the context of other measures of metacognitive sensitivity that exist? (meta-d', AUROC (Type 2), etc.)

(7) Page 8 - Another one of the results on page 8 is worth reflecting further upon: the authors note how in Experiment 1, no credible difference was found between unimodal and bimodal trials (DeltaM = -0.25 [-0.59, 0.10]), but in Experiment 2, "we observed higher metacognitive efficiency in unimodal compared to bimodal trials (DeltaM = -0.28 [-0.54, -0.02]. Those DeltaM values are nearly identical, so without a power analysis motivating the number of participants the authors collected, how certain are they that the results from these two experiments are really that distinct? It reminds me a bit of the Andrew Gelman blog post, "The difference between significance and non-significance is not significant".

(8) Is there any way to look at whether the presence of multisensory hallucinations (or perhaps that word is too strong, and we should simply consider them miscategorizations) increased as the task progressed? That is, the authors have repeated presentations of audiovisual stimuli for at least some percentage of the trials. Since the percentages for auditory stimuli being correctly categorized as auditory are at 85% in Experiment 1 and 79% in Experiment 2, were the trials where they miscategorized these stimuli equally spread throughout the task? Or did they come later in the experiment, after being repeatedly exposed to multisensory trials?

(9) Would the authors obtain the same results if they got rid of the amodal confidence judgment in their task, and simply had participants report the bimodal confidence following the presence/absence judgment? Part of the reason for asking this is that, according to page 11, the model is only fitted to amodal detection accuracy and response time data. This surprised me. I would have expected that the bimodal confidence would provide more useful information for the model fit. The authors should further explain this rationale in the paper. It seems odd to me to have the multisensory confidence ratings and not have them play a central role in the modeling work.

(10) In Figure 6, it appears the model is a bit off in its estimate of auditory responses (panel B, E) in the AV condition. Do the authors have any intuitions about why this might be happening?

(11) The authors talk about how the model is reproducing effects in the human data, but there's no systematic comparison, quantitatively, of how the two things relate. The authors should include some quantitative measure that reflects this.

(12) Related to this, I am not sure I agree with the characterization in Figure 7 that "when confidence followed a disjunctive rule, the model failed to capture important aspects of the data. On the other hand, when confidence followed a conjunctive rule, it reproduced confidence in presence judgments but failed to capture variability in confidence ratings for absence judgments." What, quantitatively, is the basis of this claim? This applies to Figure 8, too. I am not clear how, specifically, and quantitatively, the authors are justifying their claims about model fits. I don't think the confidence asymmetry index in Figure 8 is enough to quantify the quality of the model fitting procedure.

(13) Is there any chance the higher metacognitive efficiency for auditory trials is simply driven by differences in the d' values across the modalities? It might be good to probe this effect further.

(14) Lastly, I think it would be interesting to look at how instructions about modality-specific attention could modulate these findings, in terms of how unimodal (unimodal visual, unimodal auditory) or bimodal attention might modulate these effects. This is an idea for future work.

This source reminds me of an experience I had over winter break while visiting family out of state. My parents and I had dinner with a family we hadn't seen since high school; their son had been one of my close friends, but I had not seen them in nearly seven years. I now relate very differently to my parents than I did as a teenager, and both my friend and I have changed significantly since graduation. The interaction felt extremely uncomfortable and difficult to navigate. I felt caught between the version of myself I present around my parents and the version tied to that older friendship, which makes the experience feel like a really good example of context collapse.

This reminds me of what I learned in my linguistics class last quarter about style shifting and code switching. Everyone does it to some extent to show belonging, community, and connection to a certain group. We switch the way we speak to project a specific image of ourselves, or we switch according to who's listening to be more like them or more different than them to establish connection or disconnect.

This section reminds me of the strong force of anonymity, and a quote I heard from somone - I think Mike Tyson. It was something about how anonymity on the internet allows people to say things that would normally get them a punch in the face. I think it captures the key aspects of anonmity. Although it could allow people to have true, genuine conversations with strangers, it also opens up the realm of trolling for people, which can do a significant amount of harm. I don't think it is black or white on if anonmity is good or not, as it really does depend on the use case.

I think it's a very interesting phenomenon that people post pictures chasing after social rewards without thinking about the ethics of it. Even if people do think about the ethics of it, they may decide that the social rewards are far higher than their moral obligations. This reminds me of when Logan Paul decided to post a video of a deceased person in the woods of Japan, completely disrespecting that person and all subjecting viewers to traumatizing content. In my opinion, any normal person would find that inappropriate to post, but I think at the time, Logan Paul must have thought the social rewards were higher than his moral obligations.

This reminds me of TikTok, where the vertical swipe is a really strong affordance. When I use it, I don’t really stop to think about what to do next and just keep swiping up without even realizing it. It feels very natural, like the app is kind of guiding me. I notice something similar with Facebook too, where the endless scrolling feed makes me want to keep going to see more posts. Honestly, I think this design makes both apps really engaging, but at the same time, I sometimes catch myself spending way more time on them than I planned because it’s just so easy to keep scrolling.

This reminds me of TikTok, where the vertical swipe is a really strong affordance. When I use it, I don’t really stop to think about what to do next and just keep swiping up without even realizing it. It feels very natural, like the app is kind of guiding me. I notice something similar with Facebook too, where the endless scrolling feed makes me want to keep going to see more posts. Honestly, I think this design makes both apps really engaging, but at the same time, I sometimes catch myself spending way more time on them than I planned because it’s just so easy to keep scrolling.

I find in interesting how the social media that we have today is similar to the early social media websites, email reminds me of texting or direct messaging and bulletin board systems remind me of posts or reposts.

I found the descriptions/explanations of asymmetry so far a bit confusing...Is it an asymmetry in power? Or it could be understood as difference in a given aspect measured by a hierarchical system? This also reminds me of how Axel Michaels explained that hybridity could simultaneously describe the oppressing and the oppressed, between which an asymmetry exists. (p.4)

This shows how injustice spreads. It reminds me of how small problems in my community can grow if ignored.

On 2022-05-03 09:27:01, user Stefan Geier wrote:

Reminds me of (ultra) slow gastric waves (magnetoelectrogastrogram with a swallowed magnet) etc. I studied years ago in normal controls and patients with IBS or Anorexia nervosa at the Max Planck Institute of Psychiatry, Munich (HÖLZL, BRENGELMANN, PLOOG ...). Was sometimes a hard work in the basement of the MPI, and sometimes an inspiring work with engaged and very interested patients. MEC and gastric nuclei might be connected. Patients might benefit from your research.<br /> Yours Stefan Geier, Haidholzen

On 2018-11-21 20:36:16, user Gloubiboulga wrote:

Great work!

Figure 2 reminds me of this figure in this paper: https://www.ncbi.nlm.nih.go...

https://uploads.disquscdn.c...

On 2015-05-19 19:24:07, user Klaus Pawelzik wrote:

This reminds me of the work with Wolf, Geisel, Kim and Bonhoeffer,

Optimal smoothness of orientation preference maps, published in 1994. Should be cited.

this is something new that I did not consider, AI can be rapidly different and that impacts its use in our lives. It reminds me of the TedTalk video on biased views in algorithms because it correlates to AI's biases and information being different depending on which one you use.

Taoism makes me think about how I try too hard to control everything in my life, like forcing outcomes instead of letting things happen naturally. The idea of going with the flow, like water, reminds me that sometimes doing less actually works better than doing more. For me, this means I should relax more and not stress over things I can’t control. If I follow this, I think I could feel calmer and make better decisions.

I like how this brings it back to real life. It kinda reminds me that we didnt learn language in a super formal way we just picked it up by being around it and using it. The example of Québec makes it feel relatable too like even just hearing a language in everyday situations can help you start learning without even trying that hard.

This reminds me of God’s command to humans. When He gave us dominion over the earth, He did not give us the rights to do whatever we want, but to care and maintain(stewarding) the earth.

This reminds me of C.S. Lewis' Hross take on the full experience.

I like how this shifts things in a more positive direction. It kinda reminds me that the way we think about language actually matters, like if we value different languages instead of judging them, it can make people feel more accepted.

The sentence “all data is a simplification of reality” really stood out to me. I like this point because it reminds us that data is never just a perfect copy of the real world. The apple example was simple, but it clearly showed that counting something as “one” can hide important differences. I think this also connects strongly to the Twitter bot example, because the result depends a lot on how people define words like “user” or “spam bot.” This made me realize that when we look at data, we should not only ask whether it is correct, but also ask what has been simplified or left out.

I think this is a reason to be skeptical of all statistics that we see or hear. This actually reminds me of all the statistics about how much water AI actually uses, and many of those statistics and numbers are fudged and different because people are measuring things in different ways, like what counts as water usage etc. So I think that its important to take statistics like that with a grain of salt because we never know what is being simplified, and we don't know the truth of the data for ourselves.

This reminds of of 1984 by George Orwell, where the government brainwashed everyone into believing that 2+2=5 and that is the objective truth. But this article makes me think that 2+2=4 may be the false objective truth that we were all brain washed to believe and that we aren't opening our minds to the subjective truth of reality.

That is the right question to ask. This reminds me of the question of who invents what, and how it is made. Were the meanings crafted by accident, or through careful development and intelligence?

The article explains that this women was faking app store ratings and downloads. This reminds me of how bots manipulate ratings, but in this case it’s being done manually in real life. I find it surprising that she would go so far and buy 100+ iphones just to do this.

I found this really interesting. Although I was young at the time this film was released, so I wasn't thinking about botting on social media, I never would have imagined that over half of the online hate for it was not "authentic." I wonder what the socio-political benefits would have been for the author(s). Why put all that man power into trolling/hating a star wars films. I also found it interesting that Russian trolls could have played a major role in it, and that brings the same question: Why? The article highlights it could be to sway people's values, or atleast sway their values against those presented in the film. Overall, this is extremley interesting and could highlight alot about the social media sphere (especially with dog piling, etc, as it could be mostly bots doing it, which reminds me of dead internet theory).

This reminds me of an article I read sophmore year in AVID. We annotated an article regarding fixed/growth mindsets. It was rather similar to this one just with less logos to back it up.

This property kind of reminds me of the refraction illusion that I was taught in middle school. Where you put a pencil in a cup of water, you could see the pencil "bending," but the light was making this illusion happen. In this case the light is traveling differently, giving us the illusion that light is "bending."

On 2020-08-22 20:39:16, user Abhishek Singh wrote:

Reminds me of one of the paper we had written before COVID - https://arxiv.org/abs/1912....<br /> This looked at the physician notes instead of X-ray. A combined multi-task learning model could be more powerful.

On 2021-09-30 19:29:51, user Jakob Heitz wrote:

Did you account for the previous infected who died or never recovered, because of long covid?

This reminds me of a study the navy did of war planes in world war 2. They found the majority of bullet holes in returned planes were in the wing tips and central body. Therefore, they decided to reinforce those parts. What they forgot was that the planes that got shot in the engines never returned to base.

On 2025-11-10 10:25:43, user Karl Morten wrote:

This is intriguing. Are ME/CFS patients better at controlling endogenous viruses/ better at clearing them? It reminds me of Jo Elson's work showing two cohorts of ME/CFS patients have less deleterious mtDNA variants compared to controls https://www.nature.com/articles/s41598-019-39060-1 <br /> Are we looking at some sort of enhanced/different response ? Mildly deleterious mtDNA in healthy controls might better prime an innate immune response ( work of Phillip West and others)

On 2020-03-26 18:01:31, user j2hess wrote:

The point is controlled rate of spread. This on-again off-again proposal reminds me uncomfortably of the sawtooth dynamcs of a predator-prey relationship. (The rabbits breed, coyote population grows until there are too many for the food supply, so there's a population crash of rabbits first and coyotes next.) There are other ways.

A British research group modeled the growth rate using graph theory. We're not a single uniformly-connected population; there are clusters of dense connections linked by fewer connections, and spread within is faster than spread without. The power law that results is a better fit than the standard exponential growth model.

So perhaps rather than on/off, you open up retail service businesses - hairdressers, coffee shops. You don't open the big venues - concerts, major league sports, mega-conferences. This provides a somewhat controlled spread of the virus, a more stable social environment, and less economic stress.