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DOI: 10.1007/s11064-024-04150-8

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DOI: 10.1007/s11064-024-04150-8

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DOI: 10.1007/s11064-024-04150-8

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DOI: 10.1007/s11064-024-04150-8

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DOI: 10.1007/s11064-024-04150-8

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DOI: 10.1007/s11064-024-04150-8

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DOI: 10.1007/s11064-024-04150-8

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DOI: 10.1007/s11064-024-04150-8

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DOI: 10.1007/s11064-024-04150-8

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DOI: 10.1007/s11064-024-04150-8

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DOI: 10.1007/s11064-024-04150-8

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DOI: 10.1007/s11064-024-04150-8

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DOI: 10.1007/s11064-024-04150-8

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DOI: 10.1007/s11064-024-04150-8

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DOI: 10.1007/s11064-024-04150-8

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DOI: 10.1007/s11064-024-04150-8

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DOI: 10.1007/s11064-024-04150-8

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DOI: 10.1007/s11064-024-04150-8

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DOI: 10.1007/s11064-024-04150-8

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DOI: 10.1007/s11064-024-04150-8

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DOI: 10.1007/s11064-024-04150-8

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DOI: 10.1007/s11064-024-04150-8

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DOI: 10.1007/s11064-024-04150-8

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DOI: 10.1007/s11064-024-04150-8

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DOI: 10.1007/s11064-024-04150-8

Resource: ImageJ (RRID:SCR_003070)

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DOI: 10.1007/s11064-024-04150-8

Resource: ImageJ (RRID:SCR_003070)

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DOI: 10.1007/s11064-024-04150-8

Resource: RRID:MGI:2160272

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DOI: 10.1186/s13071-024-06266-w

Resource: (BioLegend Cat# 307629, RRID:AB_893575)

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DOI: 10.1186/s13071-024-06266-w

Resource: (BioLegend Cat# 348801, RRID:AB_10612570)

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DOI: 10.1186/s13071-024-06266-w

Resource: (BioLegend Cat# 318322, RRID:AB_893389)

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DOI: 10.1186/s13071-024-06266-w

Resource: (BioLegend Cat# 300406, RRID:AB_314060)

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DOI: 10.1186/s13071-024-06266-w

Resource: AB_2637396

Curator: @scibot

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What is this?

DOI: 10.1186/s13071-024-06266-w

Resource: (Thermo Fisher Scientific Cat# 25-0149-42, RRID:AB_1582276)

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DOI: 10.1186/s13071-024-06266-w

Resource: (Proteintech Cat# SA00001-2, RRID:AB_2722564)

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DOI: 10.1186/s13071-024-06266-w

Resource: (Proteintech Cat# 10494-1-AP, RRID:AB_2263076)

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DOI: 10.1186/s13071-024-06266-w

Resource: (Bioss Cat# bs-0086R, RRID:AB_10856457)

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DOI: 10.1186/s13071-024-06266-w

Resource: (Bioss Cat# bs-0479R, RRID:AB_10855328)

Curator: @scibot

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What is this?

DOI: 10.1007/s00259-024-06722-w

Resource: RRID:Addgene_48

Curator: @scibot

SciCrunch record: RRID:Addgene_48

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What is this?

DOI: 10.1007/s00259-024-06722-w

Resource: RRID:Addgene_12259

Curator: @scibot

SciCrunch record: RRID:Addgene_12259

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DOI: 10.1007/s00259-024-06722-w

Resource: RRID:Addgene_12253

Curator: @scibot

SciCrunch record: RRID:Addgene_12253

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What is this?

DOI: 10.1007/s00259-024-06722-w

Resource: RRID:Addgene_12251

Curator: @scibot

SciCrunch record: RRID:Addgene_12251

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What is this?

DOI: 10.1007/s00259-024-06722-w

Resource: RRID:Addgene_48688

Curator: @scibot

SciCrunch record: RRID:Addgene_48688

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What is this?

DOI: 10.1007/s00259-024-06722-w

Resource: RRID:Addgene_74279

Curator: @scibot

SciCrunch record: RRID:Addgene_74279

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What is this?

DOI: 10.3389/fimmu.2024.1355130

Resource: ModFit LT (RRID:SCR_016106)

Curator: @evieth

SciCrunch record: RRID:SCR_016106

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What is this?

DOI: 10.7554/eLife.91141

Resource: AB_2286802

Curator: @evieth

SciCrunch record: RRID:AB_2286802

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What is this?

DOI: 10.1016/j.isci.2023.108545

Resource: (BDSC Cat# 5072,RRID:BDSC_5072)

Curator: @maulamb

SciCrunch record: RRID:BDSC_5072

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DOI: 10.1016/j.isci.2023.108545

Resource: RRID:BDSC_5841

Curator: @maulamb

SciCrunch record: RRID:BDSC_5841

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What is this?

DOI: 10.1016/j.isci.2023.108545

Resource: RRID:BDSC_52309

Curator: @maulamb

SciCrunch record: RRID:BDSC_52309

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What is this?

DOI: 10.1016/j.isci.2023.108545

Resource: (BDSC Cat# 59819,RRID:BDSC_59819)

Curator: @maulamb

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DOI: 10.1016/j.isci.2023.108545

Resource: (BDSC Cat# 869,RRID:BDSC_869)

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DOI: 10.1016/j.isci.2023.108545

Resource: (BDSC Cat# 4781,RRID:BDSC_4781)

Curator: @maulamb

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What is this?

David Lynch Interview supposedly... source? (asking mrtnj at https://discord.com/channels/992400632390615070/992400632776507447)

Interesting with respect to orality almost more than literacy.

Sustainability reporting driven by financial materiality and promoted by, for example, the accountancy community through (e.g., IFRS Foundation) might have an insignificant or negative impact on sustainable development

• The Annals of the University of Oradea, Economic Sciences TOM XXXI, 2nd Issue, December 2022 109 / WHO IS GOING TO WIN: THE EU ESG REGULATION OR THE REST OF THE WORLD? – A CRITICAL REVIEW

—David Lynch

This is from his book: see: https://hypothes.is/a/Swp61GITEe6dmD-RxxFY4w

Why would one "leave" a link live? This statement assumes the link is live by default. A fully justified URL violates WCAG SC 2.4.4 (https://www.w3.org/WAI/WCAG21/Understanding/link-purpose-in-context.html). DOI links, as URLs, should be unlinked or violate another, likely more important, Web standard.

Wrong! DOIs should not be presented as live links because it violates WCAG SC 2.4.4 (https://www.w3.org/WAI/WCAG21/Understanding/link-purpose-in-context.html) or does APA not care about other standards?

In conclusion, the Regan artifact was initially evaluated with the neo-Aristotelian method focused on the units of analysis ethos and delivery. A deeper investigated was conducted using the situational method outlined by Bitzer, along with generic criticism supported with secondary sources by Hart and Gustainis. In this tragedy address there was the exigence, the Challenger exploding, the audience, the American people, and the constraints of only the audience being able to respond emotionally by possibly having hope in the NASA Space Program again. Tragedy address’ and crisis rhetoric that were present in the Regan artifact were also in the other speeches analyzed - Hurricane Katrina and the Pearl Harbor attacks. In all three speeches, the President(s) were addressing the nation over a shared tragedy, calming crisis emotions, and restoring hope to broken country. All three situational and all three falling into the same gerne of tragedy address. As evident in theses speeches and in other crisis rhetoric, the speaker must stay ahead of the blame and often, has to divert the nation from the truth.

A fine example of Regan implementing the methods of situational rhetoric. He is listing other similar situations of tragedy in the face of exploration that required certain rhetoric for progress, exigence, audience, and constraint.

The situation creates the rhetoric, rhetoric is a kind of force and it's supposed to change what people think. The speech is the rhetor's interpretation of reality, communicated to an audience, and that’s how knowledge, or in the case of my artifacts, hope, is created. The audience(s) were able to respond to the situation emotionally due to the rhetors forward-thinking and tone.

Another piece of evidence that displayed that Regan’s ethos was a successful means of persuasion in rebuilding faith and restoring trust in the NASA Space program was when Regan stated, “I’ve always had great faith in and respect for our space program”. The implication here was that if Regan, a man of solid ethos, still respects and trusts in the program, then so should the rest of the nation. His charismatic character encouraged Americans to follow his leadership, take his words of mourning to heart, and have faith in the NASA Space Program again. Regan was putting his credibility on the line when he said that he had great faith in the program because there have been disasters before (his reference to the explosion on the ground), and that there would possibly be more disasters in the future. Regan was bracing the public and they should trust him because he was a good man who believed in paying the price for freedom, “that’s the way freedom is, and we wouldn’t change it for a minute.” Good men and women fight for freedom. Good men and woman die for freedom. And let it be known, good men and women will continue to fight for freedom in the face of tragedy. For America to continue to enjoy the free world and trust in the leader of the free world, the nation needed to trust the NASA Space Program again.

J. Justin Gustainis, author from the article, “The Generic Criticism of Social Movement Rhetoric” from the journal, Rhetoric Society Quarterly (1982), says “genres have been defined by similarities in audience, by similarities in modes of thinking, by similarities in situations,” (252). When comparing the similar generic methods of the Regan address, Pearl Harbor, and the Bush speech, we can see that the audiences were in similar emotional crisis with the similar situation of national disaster. With situational criticism and this genre of speech, tragedy address, the problem first arises and then the rhetoric to soothe and give hope to the audience.

In this sentence, Regan uses his ethos and delivery found in the neo-Aristotelian method to encourage Americans to move forward, have faith, and trust the NASA Space Program again.

http://proxy.wexler.hunter.cuny.edu/login?url=https://www.jstor.org/stable/3885165

I believe that the rhetor, President Regan, had a fitting response to the situation addressed within this artifact. The situation at hand was a part of exploring and discovery, as in, this type of tragedy can happen when you explore the unknown. One situation, the desire for space exploration, creates a tragedy situation because of human error and pressure. The audience is limited in their actions because it’s the majority of the American population that witnessed the explosion and is in emotional crisis due to this situation. This collective situation of mourning, Regan must begin the healing process with his words of hope. The only action that the audience can take was to continue to trust the NASA space program in light of this tragedy. To have faith again in space exploration even though wounded now. President Regan achieved his goal of consoling Americans and concealing the cause of the crisis, by rhetorically framing the situation as status quo.

Here is a fine example of an observed strategic patterns found in many tragedy address’ or crisis speeches that help constitute the artifact being analyzed with a generic criticism lens. When the audience is connected to the disaster or crisis in an emotional way, the rhetor must stay ahead of the questions of blame. Who is the blame for this tragedy? “No one” says the rhetor, “these things happen,”. Sonja K. Foss says in Rhetorical Criticism Exploration and Practice that "genres influence you to develop your message in particular ways - they serve as prescriptive, ready-made patterns of communication that you can use as templates," (180). This offensive verbiage, as in staying out front of the questions before they arise, will be found as a ready-made strategy in the genre of tragedy address.

The best visual evidence that suggests Regan was successful in his use of delivery was when he said, “give me a challenge, and I’ll meet it with joy”. At 1:36 of his speech as Regan said “give”, his brow lightens, his eyes lift, and a small smile emerges on his face for the duration of this quote, and then immediately returned to the somber furrowed brow look. Regan was a trained actor and was effectively able to change and move through emotions and facial expressions long before he was a U.S. President. The ability to have earnest expressions and show compelling emotions makes it easier to persuade an audience to the rhetor’s favor and heed his message. When Regan lifts his brow and smiles ever so strategically, it gave the audience hope and hope leads way to trust, and trust leads way to unity and faith in NASA Space Program. In this kind of tragic moment, if one smiles too long or too wide, it looks inappropriate or staged. The entire dynamic of the speech was mourning a tragedy, but at 1:36 Regan found a way to insert a dose of joy, and finding an appropriate dose of joy in this terrible moment was needed by all. Regan spoon-fed much needed hope right to millions of Americans. The smile was calculated with the precise word of “joy” to have an impact and the lasting effect was hope and faith restored in NASA.

National disasters like Pearl Harbor, September 11th, COVID -19, all required situational rhetoric, with the existence of the exigence, audience, and constraint, but also the speeches connected to these events fall into genre of crisis rhetoric as well.

Here is an example of another address of a national tragedy by the U.S. President Franklin D. Roosevelt on December 7th 1941. The exigence was the attack on Pearl Harbor, the audience was the American people, and the constraint was the fear of more lives of Americans lost at war. Again, we see a U.S. president taking situational rhetoric to address a national tragedy and implementing frame-works of similar speeches of the same genre. The president must stay ahead of the audience and press and at the same time give hope of victory to the nation.

https://www.youtube.com/watch?v=pU1HycD_0p4

For example, another national tragedy that required the genre of crisis rhetoric was President George Bush’s address regarding Hurricane Katrina.

With the genre of tragedy address or crisis rhetoric there is the tragedy, often a natural disaster like a hurricane. The connective content in these types of speeches is a reassuring patriarchal tone of a father or a U.S. president. A common organizing principle of both the Regan address and the Bush address was to give hope. The rhetor must be able to give the audience hope of a better time. Both presidents after they laid out what happened and why, shifted their tone and words to that of uplifting hope for the nation.

Here is U.S. President George W. Bush in the Rose Garden on September 5th addressing the AP about the national disaster, Hurricane Katrina. The viewer will note that towards the end of both short tragedy address (Regan's and Bush's), the president's tone shifts towards hope, grace, and uplifting Americans.

https://youtu.be/P70HllXsz1c?si=sCwDfEFqWVK6ASZK

The units of analysis for generic criticism are artifacts having a situation in common, like a tragedy or crisis. There must be substantive and stylistic features within the content of that genre of rhetoric that are similar and comparable to one another. One must be able to analyze the common traits found in tragedy address or crisis rhetoric. Finally, there must be an organizing principle to qualify as a generic method. In this case, the organizing principle is a national crisis with the space shuttle challenger exploding in front of the American people, deeply traumatizing them. Mourning this monumental loss as a nation was a crisis.

Authors Paul ‘t Hart and Karen Tindall in their book chapter entitled, “Understanding Crisis Exploitation: Leadership, Rhetoric and Framing Contests in Response to the Economic Meltdown” from the book, Framing the Global Economic Meltdown, say about the genre of tragedy address and crisis rhetoric is that, “in today’s risk society, disasters typically evoke nagging questions that spell trouble for incumbent leaders: why did they not see this coming?” (25). The content of both addresses must acknowledge upfront that Americans were going to be upset and emotional about a tragedy like the Challenger and a disaster like Hurricane Katrina. These events caused crisis in the American population. The people in charge were going to be questioned and someone is going to have to be blamed. In crisis rhetoric and tragedy addresses, the rhetor must present with a “this has never happened before” or “it’s a cruel natural disaster that we could do nothing about” mentality. The speech must stay ahead of mass blame, and we see that in both addresses. It’s no one’s fault, and now we must unite in hope and move forward as a country.

In the case of the space shuttle challenger, the writers and producers of the Netflix original docuseries, “Challenger: The Final Flight”, speculate that NASA was aware of the faulty ring that caused the explosion and had several failed launches. One could argue that at this point in the speech, “we’ve never had a tragedy like this,” that President Regan was lying to the American people. He was using crisis rhetoric tactics of covering up the questions of how and why this could ever happened before they arose in the audience.

https://www.netflix.com/watch/81012171?trackId=255824129

https://www.jstor.org/stable/j.ctt24hf3m.4

In this introductory paragraph, the units of analysis for situational criticism are in full effect with the exigence (the problem), audience (those who must be able to change heart or take action), and constraints (factors that limit or influence the audience’s action). The problem is that the shuttle crashed, and Americans are devastated and will lose faith in the NASA space program. The audience is the American people, the same people who witnessed the tragedy earlier in the day and were listening to the President speech that same evening. The constraint is that such a terrible event occurred live on TV with so many families emotionally invested in the outcome and nothing they could do but mourn. According to Lloyd F. Bitzer in his article, “The Rhetorical Situation” from the journal, Philosophy & Rhetoric, he states regarding situational rhetoric is that “a work of rhetoric is pragmatic; it comes into existence for the sake of something beyond itself; it functions ultimately to produce action or change in the world; it preforms some task,” (3). The tragedy address is situational discourse because the speech is only happening because the shuttle exploded, otherwise the President would have been giving the State of the Union Address. The desired action of the audience is to have hope, heal, and to not lose faith in NASA.

http://proxy.wexler.hunter.cuny.edu/login?url=https://www.jstor.org/stable/40236733

Ronald Regan was the 40th U.S. President and on January 26th of the year 1986, he was slated to give the yearly State of the Union Address. However, the space shuttle Challenger had just exploded during takeoff a few hours earlier on live TV. The Challenger was set to bring the first civilian into space, a beloved teacher named, Christa McAuliffe. President Regan had to use various means of persuasion during his speech to console a crushed and heartbroken nation, as well as rebuild American’s confidence in the NASA Space Program in the face of this tragedy. This paper will analyze the canons of rhetoric, particularly the combination of the president’s ethos and his delivery, to persuade the American public into trusting the NASA Space Program again after such a national loss. Beyond the neo-Aristotelian approach, a further analysis of this artifact will use the secondary method’s, situational criticism, and generic criticism to reveal Regan’s rhetorical mastery.

Below is a video from ABC Nightly News with Lester Holt aired on January 28th, 1986 the day of the launch of the Space Shuttle Challenger. This video will give a sense of the dire situation at hand and how that required situational rhetoric. Furthermore, this video will exemplify why and how this tragedy speech can be analyzed with the generic method of criticism and placed in its own genre of speeches called crisis rhetoric. As one can view in the video, America was in crisis.

https://youtu.be/yibNEcn-4yQ?si=wbe2X8arIxYnVc0E

Network

In this article, Jacobson shares the story of her distributing experience of a cyberstalker. Her experience illustrates the profound and enduring impact of cyberstalking while also highlighting the challenges of seeking justice and protection.

It was interesting to see just how organized and discreet but also open they are in what they believe in and what their values are. One detail that stood out to me was that they don't just target black people but pretty much every other ethnic group including Asian Americans. I thought that they were exclusive to just African Americans but reading that they weren't was new to me.

"When it comes to Twitter — or to Facebook, or Instagram, or Snapchat — blocking is relatively uncontroversial." The article discusses about the needs of blocking features. However, I wonder how affective are these blocking features as there are more complexity to it and its hard to track online activities.

I believe doxing is a very bad consequence of our digital world. It involves publicly sharing someone's personal information online without their consent, often leading to severe consequences for the victim. This information can come from public databases, social media, or even private details obtained through hacking. While some forms of data aggregation might be legal, they can still be associated with stalking and intimidation. Doxing is often used for malicious purposes like online shaming, extortion, or vigilante actions. This harmful practice highlights the darker side of our connected world and the need for stronger protections against such invasions of privacy.

It is my opinion that the "cancel culture" contributes to the fact that social media in this day and age is so sensitive. When individuals on the internet find something unpleasant and detest it, they have the ability to ruin a person's career via the phenomenon known as cancel culture. Although cancel culture may be beneficial in that it helps to safeguard and make the internet a safer place, it also results in a great deal of limitations that are not essential.

I believe crowd harassment is unjustifiable, it’s not an appropriate or ethical way it only perpetuates fear and hostility, undermining any chance for constructive dialogue or resolution.

It is interesting and at the same time sad to see just how much hate and misinformation can spread from such a small group of people that try to misconstrued someone's life. I relate to this as rumors that usually spread either at school or within clubs amongst friends and people are usually rooted in a small group or singular individuals who don't like someone.

Group harassment is often more serious than individual harassment because it is not just the malicious behavior of one person, but a joint attack by many. This phenomenon is very common on the Internet and has even led to the formation of the so-called "cyber violence". I personally believe that social media platforms and the government should work together to formulate tougher laws and regulations to curb the spread of this phenomenon. At the same time, users themselves need to improve their online literacy to avoid participating in or contributing to online violence.

Teste ok

teste

The argument/ideology that pins down Barthes’ deconstruction of the Eiffel Tower is very Nietzchian. Much like Nietzsche’s popular argument that art is the only truth because it allows one to live in a personal abstraction and intuition, the tower being art means it surpasses our rationalization, deconstruction, and assimilation of it into one side of binary schemas. It exists to emphasize its inability to be known by us and to serve almost “mythical” purposes that transcend rational rules of the world. In other words, “Barthes’ phenomenological approach brings us to the focus of our investigation: an architectural structure’s capacity to simultaneously be understood as agent and object, a capacity we regard as a peculiar oscillation between function and symbol in the case of the Eiffel Tower” (Steiner).

There is a lot to unpack with the contradictory qualities of the “utterly useless monument”, which we actually learn is pretty useful (Barthes 5). The point that stands is that, physically, the tower is an uncontainable object that we try to domesticate. One way we do this is through “the installation of a restaurant [...or other] means of leisure” in the tower itself (Barthes 16). The fact that the tower is an open construction makes us uncomfortable when we are used to typical tourist hotspots (like museums, for instance) being enclosed for us to feel like we entered, experienced, and “owned” some of it. The tower doesn’t do that for us. So, we have to create a mini world surrounding the tower in order to make it feel normal. In our conception of the order of the world, the Eiffel tower is unique to us because it is simultaneously a representation of the inside and of the outside world. This quality, that the tower is somehow both sides of an opposite binary, is too far outside of the social contract, and Nietzche would say (and Barthes points to it) that we often try to tackle this discomfort by trying to reduce the tower. We do this by turning the tower into a sight of projection. It becomes a symbol of industrialism, of Paris, of travel, of art, of Paris itself–whatever one may choose. But it is in this choice, that we strip the tower of the other symbols it projects equally as strongly. And this is where the problem lies. We must look at the tower as the embodiment of all the opposites it may be: inside/outside, industry/art, ugly/beautiful, all at the same time.

Barthes asks us to consider why the tower makes us so uncomfortable in this binary presentation. Perhaps it is because this makes the tower oddly more powerful than us. The tower can be a spectacle and an object, useless and useful, inside and outside. We cannot be those things. If we are looking at the tower, we can't be in it, for example; but the tower can be both an empty base space outside, and an indoor restaurant as well, for example. None of our relations to the tower can come together at the same time, while the tower can be opposites at the same time. We can only perceive the tower as one of its opposite meanings at a time, and we have to kind of deal with the impossibility of bringing together two things that are true and simultaneous but also cannot co occur logically. I think one way we do this is by glossing over it all and pretending everything can occur at the same time–a comforting thought facilitated by the constructed surrounding environment.

However, by doing this, what simultaneously happens is that the tower becomes a signifier of basically an infinite sight of projection. It is reduced to a symbol of Paris, of travel, of industrialism, of some kind of focal point in France. The tower being a signifier for everything really just makes it nothing. And when we come face-to-face with this (structural and symbolic) emptiness, we rush to find ways to create more perceived “somethingness”(we add restaurants, shops, carts of food, and other community experiences all around the tower) to fit into our schemas and orders.

Barthes, Roland. The Eiffel Tower - Roland Barthes - LANTB, lantb.net/uebersicht/wp-pdf/eiffelTower.pdf. Accessed 13 May 2024. Steiner, Henriette, and Kristin Veel. “Towering invisibilities: A cultural-theoretical reading of the Eiffel Tower and the One World Trade Center.” Qualitative Inquiry, vol. 25, no. 4, 5 Aug. 2018, pp. 407–416, https://doi.org/10.1177/1077800418790297.

When picking one element of Pentadic Criticism to deconstruct the Eiffel Tower with, "scene" seems to be an influential factor: would the Eiffel Tower have become such a core part of identity, or have been so controversial, in any other city? For this we might look to the impact of monuments such as One World Trade Center, or the Leaning Tower of Piza, and compare and contrast features of the artifact. I would argue that the Eiffel Tower is only as influential as it is, and as core to the representation/identity of Paris BECAUSE it is in Paris. Its design was so different from anything popular during the time it was built, and this initial legacy seems to carry on in different interpretations throughout the years. It has turned into a display of the importance of engineering, industry and development, and turned into a display of...well, as explained through the feminist and ideological critiques... just about anything!

For the end: an excerpt from a research journal that I could not have stated better myself: "The Tower has been pro- claimed the symbol of industrial and artistic progress, the metro, electric lights, elevators, telephones, military power, centralization, the union of workers and engineers, mathematical energy, technical utopianism, mod- ern Paris, practical science, industrial and artistic progress, the union of workers and engineers, superhuman exaltation, the marriage of skill and imagination, and architectural eclecticism. It has been described as a gra- cious palm tree, mast, aging trunk, giraffe, monument and poem, tall and skinny pyramid, tragic lamppost, disgraceful skeleton, ancient god and modern beast, colossal golden Phare of industry and capital, and modern Tower of Babel. To quote Cendrars again: "Tu es tout / Tour." For Parisians it is the inevitable physical presence, while for the rest of the world it is the inevitable symbol of France"

Thompson, William. “‘The Symbol of Paris’: Writing the Eiffel Tower.” The French Review, vol. 73, no. 6, 2000, pp. 1130–40. JSTOR, http://www.jstor.org/stable/399367. Accessed 14 May 2024.

It is notable that the Eiffel Tower is made of iron. Iron as the building material makes the Eiffel Tower made by man, for man, and of man. This changes how the Eiffel Tower might be seen or interpreted on the surface. In fact, it might be the very opposite of what we assume the tower to be on the surface. The Eiffel Tower is most commonly the symbol for Paris–the city of love. Just as Mars is the god of war and symbol of man, love is the symbol for women, belonging to the goddess Venus (Venus). Thus what we imagine the Eiffel Tower to be, is actually not what it is at all. It is not a feminine representation of love, but rather a towering and tough masculine force.

pentadic criticism can lead to questions like, how well did the agent/artifact use the available means to achieve their goal? With the Eiffel Tower, one might say that Gustave Eiffel is not even the agent, rather the tower itself is. And so, the tower is successful in highlighting the importance of mechanical ingenuity as well as artistic ingenuity, because it has become a universal symbol for Paris.

Pentadic criticism can be used to analyze the Eiffel Tower as well. It requires of us that we identify 5 items of the pentad: * Agent (who is performing the act) * Act (what is happening) * Scene (where and when the artifact was produced) * Purpose (why) * Agency (how/what means does the agent use) Pentadic criticism allows us to assign various different characteristics or details to each “item”, resulting in various interpretations of the same artifact. (Foss 356) Here is one possible pentad and interpretation of the Eiffel Tower * Agent: Gustave Eiffel * Act: constructing the Eiffel Tower * Scene: Paris during 1889 * Purpose: to introduce the value of engineers as creative artisans and mathematical intellectuals in a climate heavily dominated by artists only Agency: by submitting the design for the Eiffel Tower to the World Fair contest, Eiffel found a means through which he could gain extreme popularity for his cause. In construction of the tower itself, the use of metal and open structures contribute to the “engineer” aspect of the monument. Next is to analyze the artifact using some combination of these five characteristics. For instance, one might argue that agency and scene are the most important qualities of this artifact: By using the World Fair contest as a way to bring attention to his industrial artifact, Gustave Eiffel also shattered a perception of Paris as a classy and elegant city. He took attention away from the belle epoque and forced people to think of how structures could also serve useful purposes. The message behind the Eiffel Tower may not have come across this way had Eiffel, an engineer, not submitted his work to an art competition like this one.

Setting is an important element not only in pentadic criticm but provides context for a feminst critique of the Eiffel Tower as well. The Eiffel Tower is 1,083ft tall (The Official Eiffel Tower). When it was first built, it was the tallest structure in the world, briefly, until the Empire State Building was erected. Regardless, such a looming force stuck out somewhat awkwardly amidst Parisian landscapes in the 1880s, which was in the middle of their “Belle Époque”, or “beautiful era”. This era extended beyond the popular and affluent-looking terracotta and cream colored architecture, but also emphasized regional peace and prosperity, and infiltrated social practices by emphasizing female domesticity (Christiansen, 2015). Here is a link to a video that provides more context about what was going on during the Belle Epoque, the values in society, and pushes towards equality that were made. https://www.youtube.com/watch?v=AK11U_AZCyE ](

Christiansen, Hope. “Grappling with feminism in the Belle époque: Colette Yver’s princesses de science and Les Dames du Palais.” MLN, vol. 130, no. 4, Sept. 2015, pp. 946–962, https://doi.org/10.1353/mln.2015.0054. “Time-Travelling Back to France’s ‘Belle Époque’ • France 24 English.” YouTube, YouTube, 23 Mar. 2023, www.youtube.com/watch?v=AK11U_AZCyE. “15 Essential Things to Know about the Eiffel Tower.” La Tour Eiffel, The OFFICIAL Eiffel Tower website, 27 Dec. 2023, www.toureiffel.paris/en/news/history-and-culture/15-essential-things-know-about-eiffel-tower#:~:text=For%20130%20years%2C%20the%20Eiffel,know%2Dhow%20and%20industrial%20genius.).

Background information on why the Eiffel Tower was built: When Gustave Eiffel started construction on the Eiffel Tower for the 1899 World Exhibition in Paris, it was easy to see the controversy in the fact that an engineer such as himself had won an art competition. Architects, designers, and everyday Parisians especially despised the tower for not fitting into the mold of what Parisian structures should look like. Rather than being constructed of stone, elegantly curated to nourish Parisian lavishness, the tower was a cold, bare slab of iron sprung up right in the middle of the city; it seemed unfathomable that Eiffel had somehow won Paris’ most noble art competition with this “glorified lamppost” (Sociéte d'Exploitation de la tour Eiffel) (Vukelj 2024)

Above almost all else for a feminist critique, we must look at the Eiffel Tower as a phallic symbol. The ideology of such a critique is rooted in the idea that any and all space has gender. Gender can be assigned “intentionally according to the sex of the architect, interpretive[ly] through theory, criticism and history, and through use (public toilets sexed male or female while the domestic kitchen is gendered feminine)” (Niculae). I have already discussed how building materials might associate gender, but another interpretation could come from examining the height or shape of the tower. The tower looms over 1000 ft over Paris, making it impossible to deny the power relations inscribed in the space, on many levels.

Compared to the “belle epoque” buildings, the Eiffel Tower screams masculinity, and dominates physically over them due to its height. It dominates the “building to building” comparison. It does the same in the “building to observer” comparison. It is a mystical object of force, towering over us, and seems to rule over or define Paris. Similarly, men rule over and define work spaces, social customs, and many other aspects very much aligned with the tower and its message.

Niculae, R. L. (2014). Gender analogies in architecture. Journal of Research in Gender Studies, 4(1), 474-490. Retrieved from http://proxy.wexler.hunter.cuny.edu/login?url=https://www.proquest.com/scholarly-journals/gender-analogies-architecture/docview/1553511966/se-2

Through a feminist criticism perspective, we can establish new meaning for the Eiffel Tower based on the fact that it is made of iron. In classic literature, iron is deemed the “Metal of Mars” and has come to symbolize probably what is most idealized in humanity, or even more specifically, in masculinity: “strength, obstinacy, fortitude, honor, courage, sharpness (of body and mind), tenacity, and confidence in power”(Cabantchik). It does not take much in-depth analysis–rather, a brief examination of the structures of our society (we even have a superhero: “Iron Man”)–to see how these features are ones deeply rooted in our perception of what it means to be masculine. Iron’s durability as a metal is intertwined with that of a man. The metal’s association with Mars has male-coded connotations dating back to Ancient Rome, as Mars was an ancient Roman god, seen as the protector of Rome, and instilled nationality and pride in Romans for their army–likely one of the most raw and courageous in the world (Mars). With iron being the metal of Mars, the connection between manlihood, iron, and strength/courage, becomes almost inseparable.

Cabantchik, Zvi Ioav. “In search for symbolic qualities of iron: The Metal of Life.” Frontiers in Pharmacology, vol. 7, 3 Aug. 2016, https://doi.org/10.3389/fphar.2016.00220. “Mars.” Encyclopædia Britannica, Encyclopædia Britannica, inc., www.britannica.com/topic/Mars-Roman-god. Accessed 13 May 2024. “Venus.” Encyclopædia Britannica, Encyclopædia Britannica, inc., www.britannica.com/topic/Venus-goddess. Accessed 13 May 2024. “15 Essential Things to Know about the Eiffel Tower.” La Tour Eiffel, The OFFICIAL Eiffel Tower website, 27 Dec. 2023, www.toureiffel.paris/en/news/history-and-culture/15-essential-things-know-about-eiffel-tower#:~:text=For%20130%20years%2C%20the%20Eiffel,know%2Dhow%20and%20industrial%20genius.).

I was in a vintage shop about 30 minutes from downtown Los Angeles earlier in the week and the proprietor had a mostly functioning 1950 Smith-Corona Sterling for sale for a roughly equivalent US $150. (One key was disconnected, but fixable, and some keys were sticky, the ribbon was disintegrating, it was incredibly dirty, with a case in very poor condition.) The Sterling was similar to the Silent, but without some of the extra bells and whistles. She wouldn't accept an offer of $40 for it, which I thought was a reach for the dreadful condition it was in. Her reasoning was that she was sure that someone (read: a sucker) would pay the $150 for it. At a yard sale it might be worth $5. Cleaned up a bit maybe $30. In online platforms they're going for a bit more, but you're also saving yourself some level of "shoe leather" in the work of searching for the exact model you want.

I've been specifically watching this model and a few related ones for a few months, and machines of indeterminate condition (though in my experience they're usually reasonably functionable or easily fixable), like this go for about $50 on ShopGoodWill.com (as auction items). There are usually about 4-5 per week which come up as this was a popular model in the 50s. You can probably find similar prices on eBay, though sellers there usually have a little more information about the working condition. They're definitely common enough that you could easily wait for the exact color options and typeface (pica or elite) that you're looking for, and could also probably purchase two for the price he's asking (including shipping.) I've been watching for a similar mid-50s Smith-Corona Clipper with similar colors and elite type for a while and just bought one online last week for $35. Patience definitely pays off.

I would only go as high as $150 on that machine if I knew it was well functioning and had a brand new platen in the last several years. You can tell him that most of the expensive machines in the range he's asking for are all fully functioning, have been well maintained and/or recently serviced, and often have new platens, rubber rollers, and feet replaced. He'll know that this isn't the case with his and may come down in price. They're likely pricing it based on other listings they see and not pricing it based on actual sales. If it's their only machine, wait things out until they see that there aren't any takers. If it's a vintage shop, simply move on.

The Smith-Corona Silents from this time period are really spectacular and solid machines, so good luck in your search for the perfect one.

Thomas Stewart Baker (born 20 January 1934) is an English actor and writer. He played the fourth incarnation of the Doctor in the science fiction television series Doctor Who from 1974 to 1981.

https://en.wikipedia.org/wiki/Tom_Baker

Doctor Who is a British science fiction television series broadcast by the BBC since 1963.

https://en.wikipedia.org/wiki/Doctor_Who

What's the joke?

The play on words is obvious.

David John Tennant (né McDonald; born 18 April 1971) is a Scottish actor. He is best known for portraying the tenth incarnation of the Doctor in the sci-fi series Doctor Who

https://en.wikipedia.org/wiki/David_Tennant

Over the last 200 years, the pronoun whom has been on a steady decline. Despite its waning use in speech and ongoing speculation about its imminent extinction, whom still holds a spot in the English language, particularly in formal writing.

https://www.thesaurus.com/e/grammar/whom-vs-who/

You use who in questions when you ask about the name or identity of a person or group of people.

Example sentences: * Who's there? * Who is the least popular man around here? * Who do you work for?

https://www.collinsdictionary.com/dictionary/english/who

You use whom in questions when you ask about the name or identity of a person or group of people. Example sentences: * 'I want to send a telegram.'—'Fine, to whom?' * Whom did he expect to answer his phone? * 'You're too sensitive.'—'Too sensitive for whom?'

https://www.collinsdictionary.com/dictionary/english/whom

Charles, Prince of Wales is the former title of Charles III (born 1948), before his accession to the throne of the United Kingdom and the other Commonwealth realms.

https://en.wikipedia.org/wiki/Charles,Prince_of_Wales(disambiguation)

Charles III (Charles Philip Arthur George; born 14 November 1948) is King of the United Kingdom and the 14 other Commonwealth realms.

https://en.wikipedia.org/wiki/Charles_III

Joseph Ellis Pasquale (born 20 August 1961) is an English comedian, actor and television presenter.

[...]

He won the fourth series of I'm a Celebrity...Get Me Out of Here! in 2004 and participated in the eighth series of Dancing on Ice in 2013. He also hosted the revived version of the game show The Price is Right.

https://en.wikipedia.org/wiki/Joe_Pasquale

Queen are a British rock band formed in London in 1970 by Freddie Mercury (lead vocals, piano), Brian May (guitar, vocals), and Roger Taylor (drums, vocals), later joined by John Deacon (bass).

https://en.wikipedia.org/wiki/Queen_(band)

Sir Brian Harold May CBE (born 19 July 1947) is an English musician, songwriter, record producer, animal rights activist and astrophysicist. He achieved worldwide fame as the lead guitarist and backing vocalist of the rock band Queen

https://en.wikipedia.org/wiki/Brian_May

The museum has over 300 restored exhibits of garden machinery from over the last 200 years, as well as a collection of lawnmowers previously owned by famous people including Prince Charles and Princess Diana, guitarist Brian May, performer and presenter Paul O'Grady, comedian Lee Mack, and Coronation Street actress Jean Alexander, who lived in Southport for many years.

https://en.wikipedia.org/wiki/British_Lawnmower_Museum

dibber definition

a tool used for making holes in soil when planting seeds or small plants

https://dictionary.cambridge.org/dictionary/english/dibber

Trowel definition

a small tool consisting of a flat metal blade joined to a handle, used for spreading building materials such as cement

https://dictionary.cambridge.org/dictionary/english/trowel

The word "tool" could mean different things:

Tool definitions:

a piece of equipment that you use with your hands to make or repair something

OR

(offensive) an insulting word for a person who you dislike very much or who behaves very stupidly

OR

(offensive) a penis. https://dictionary.cambridge.org/dictionary/english/tool

So "Famous tools" could mean:

Equipment (tools) that are recognised by many people.

OR

Famous people who are disliked very much or who behave very stupidly.

OR

Famous penises a.k.a. dicks, dongs, knobs...

Where's the joke?

I think the audience laughed because it sounds like Lee could be talking about looking for famous penises on a website (or I just have a dirty mind).

Frederick "Fred" Flintstone is the main character of the animated sitcom The Flintstones, which aired during prime-time on ABC during the original series' run from 1960 to 1966

[...]

Fred's trademark catchphrase yell is "yabba dabba doo!"

https://en.wikipedia.org/wiki/Fred_Flintstone

Listen to how Fred says Yabba Dabba Doo! on YouTube

Where's the joke?

This catchphrase "yabba dabba doo" sounds similar to the question "What does your dibber do?".

Lee is asking Dale if he is Fred Flintstone because he sounded like him when he asked the question "What does your dibber do?"

Inspector Morse is a British detective drama television series based on a series of novels by Colin Dexter.

https://en.wikipedia.org/wiki/Inspector_Morse_(TV_series)

The British Lawnmower Museum is a museum dedicated to the history of the lawnmowers in Southport, Merseyside, northern England.

https://en.wikipedia.org/wiki/British_Lawnmower_Museum

The nation's foremost garden machinery collection, multi-award winning unique museum of over 250 restored machines.

https://www.tripadvisor.co.uk/Attraction_Review-g191255-d564490-Reviews-British_Lawnmower_Museum-Southport_Merseyside_England.html

Would I Lie to You?...

(abbreviated as WILTY) is a British comedy panel show aired on BBC One.

For each show, two celebrity guests join each of the team captains. The teams compete as each player reveals unusual facts and embarrassing personal tales for the evaluation of the opposing team. Some of these are true, some are not, and it is the panellists' task to decide which is which.

[...]

In all rounds, the scoring system is the same: teams gain a point for correctly guessing whether a statement is true or not, but if they guess incorrectly the opposing team gets a point.

https://en.wikipedia.org/wiki/Would_I_Lie_to_You%3F_(British_game_show)

'possessions' element was introduced, in which the panellist takes an item out of a box and reads a statement from a card, and has to convince the opposing team that the possession genuinely belongs to them. https://en.wikipedia.org/wiki/Would_I_Lie_to_You%3F_(British_game_show)

Good idea to promote attending smaller venues while giving an opportunity for smaller musicians to get exposure and experience. PROMOTION is KEY to breathing live into the small music scene again!

Important stat for music venues

Probability

Legal and moral reasoning

Moral reasoning

Moral reasoning

Legal reasoning

Legal reasoning

Moral reasoning

Moral reasoning

Legal (constitutional) and moral reasoning

Causality

Causality

Causality

Causality

Extended Argument

Extended Argument.

This section gives many reasons and ways the overturning of this law will impact women across the nation.

I almost feel as if this is a premise and an enthymeme. This is the title of the argument and a premise all in one, I feel as though this argument should have a different title and this should be a premise in the argument.

This is a valid argument

Fallacy: Loaded question

Cognitive Bias: Confirmation Bias

Here, and throughout the entire article, the author is only presenting points and quotes that align and agree with her own viewpoint, failing to also present and debunk any counterarguments.

Cognitive Bias: Attribution bias

Here, the author is attributing individuals being either republican or religious to explain their pro-life belief system, which, in a lot of cases, simply isnt true.

Fallacy: False dichotomy

I believe that this is and instance of false dichotomy because the author is essentially stating "you either have money to travel and get an abortion, or you are forced to carry a baby to term", which is simply not true. The author also fails to mention any alternative solutions to this issue.

The phenomenon of personal harassment on social media concerns me deeply. As technology evolves, harassers have more means to violate the privacy and safety of others. Cyberstalking and hacking, in particular, are not just psychological fears, but may pose serious threats to the real life of the victims. I believe that social media platforms need to take more responsibility to protect the safety of their users through stricter regulation and more effective reporting mechanisms.

I do agree with the statement that individual harassment has always existed, but social media added onto it. Its crazy to think how bullying, cyberstalking and tracking is more common within the social media space. It is also tricky because what you post is basically a history of your life in a way and it's all public information even if your account is private.

Individual harassment has always existed, but social media has introduced new methods for it. These include bullying, where mean messages are sent through DMs, and cyberstalking, which involves persistently finding and following someone's account. Hacking into accounts to uncover secrets or make threats is another method. Tracking involves monitoring a victim's social media to control their interactions, often using spy software. There are also severe threats like death and rape threats. These new tools make harassment easier and more pervasive, highlighting the need for stronger protections and awareness on social media platforms.

Personally, I think harassment done online is a growing issue in today's world.

There are several instances in which, once we have registered a piece of software, they will disclose our personal information, including our name and cell phone number. Scammers will be prompted to make harassing phone calls, send harassing messages, or send harassing email boxes as a result of this. The messages that are annoying are quite difficult to understand. They will solicit us for money and may even get dangerous if we take the call or trust what they are saying they are saying. Additionally, there will be occasional instances of harassing pornographic messages appearing in the messages of strangers on Instagram.

Note: This response was posted by the corresponding author to Review Commons. The content has not been altered except for formatting.

Learn more at Review Commons

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Reply to the reviewers

Dear editor and reviewers,

we thank you very much for your constructive comments, criticisms and suggestions for improvement of our manuscript. We have addressed all points raised by you and have added our point-by-point response to your comments below.

With best regards on behalf of all authors,

Andreas Wodarz

1. Point-by-point description of the revisions

Reviewer #1

Evidence, reproducibility and clarity

Baz/Par3 is an important conserved protein acting as a master regulator of cell polarity in a wide range of cell types. This study focuses on re-assessing the subcellular localisation of Baz/Par3 in a range of Drosophila tissues. This is an important study with respect to our understanding of Baz/Par3, as there have been conflicting reports on the localisation of Par complex members - while the majority show localisation to cell cortex and intercellular junctions, several reports have claimed that Par complex members localise at additional subcellular sites including the nucleus, nuclear envelope and neuromuscular junction. In this study the authors re-assess this issue for Baz/Par3 in a comprehensive and thorough manner.

We thank the reviewer for this overall positive assessment of our work.

*1. They used a variety of antibodies raised in different host animals against different epitopes of Baz 2. They tested the specificity of these antisera using mosaic analysis with null mutant baz alleles and tissue-specific RNAi against baz 3. They used a GFP-tagged Baz under control of its endogenous promoter in a baz null mutant background to compare with the subcellular localisation of the respective GFP-Baz fusion proteins to the staining results with anti-Baz antisera

The data from each of these experiments are very clear and convincing. Comprehensive methods are included which means that each of the experiments with specific anti-sera/RNAi lines/GFP-tagged conditions could be reproduced. There are a couple of experiments which were performed in support of the conclusions (extra RNAi lines and stronger expression of Gal4) listed as (data not shown). I would strongly suggest including these data as extra supplemental figures. Together, their results clearly show that Baz/Par3 localises to the cortex and intercellular junctions, but that anti-sera staining at the NMJs and nuclear envelope appear to be a staining artifact, likely due to staining with an unidentified epitope.

Minor comments 1. Many of the figures have overlays of red and green which will be indistinguishable from each other to colour-blind readers. Please alter to make colour-blind friendly (eg magenta-green)*

We have changed all figures in the following way: All single channel images have been converted to inverted grayscale to improve the visibility of weak fluorescence signals. In all multicolor overlay images, red has been omitted and instead green, magenta, blue and grayscale have been used to improve the visibility for color-blind readers.

2. In Fig 2D please indicate where the epidermis and neuroblasts are

We assume that the reviewer refers to Fig. S2D. In the revised version of the manuscript, this figure is now Fig. S2A. We have marked epidermal cells and neuroblasts by different symbols.

*3. In the following two places there are experiments describe where the data is listed as not shown. Please show the data as additional supplemental data. They are P8 - This result was confirmed using the CY2::Gal4 driver line expressed in the follicular epithelium and with three different RNAi lines against baz (data not shown). *

We have deleted this sentence because expression of CY2::Gal4 in our hands was weaker and thus the RNAi effects less reproducible than with tj::Gal4.

P11 - We also did not see any downregulation of Baz or a-spectrin upon baz-RNAi in M12 at 29°C, when the UAS-Gal4 system is maximally active (data not shown).

We now show these results in the new Fig. S8.

4. Figure 3 - this would be easier to interpret with a few arrows/arrowheads indicating the NMJs

We have added arrows pointing to NMJs and arrowheads pointing to nuclei.

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Significance

It will be important to publish these results as it means that findings for a function of Baz/Par3 at the NJM and the nuclear envelope should be regarded with caution, and it may save researchers chasing for functions for Baz/Par3 in places where they are simply not expressed. As much of our fundamental understanding of how Par3 works in vertebrates has its roots in studies in Drosophila, this is likely to be of wide relevance.

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Reviewer #2

Evidence, reproducibility and clarity

*Evidence, reproducibility and clarity

1.1 Summary

This reviewer acknowledges the expertise and contributions of Prof. Wodarz and his research group in the field of development, cell polarity regulation and Drosophila genetics.

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Manuscript summary:

Kim S. et al. explored the localisation of Bazooka, the Drosophila homolog of the polarity protein Par-3, at two non-canonical positions for a cell polarity factor: the nuclear envelope in epithelial tissues and the postsynaptic membrane of the neuromuscular junction (NMJ). Previous work has shown the detection of Par-3/Baz at the nuclear envelope and the NMJ using antibodies against Par-3/Baz. Here, the authors used a combination of genetic perturbations (baz RNAi and generation of genetic mosaics for baz) and GFP-labelled Bazooka lines to test if the antibody-mediated detection of Baz at the nuclear envelope and NMJ is artifactual. The data provided by the authors strongly suggest both the nuclear envelope and NMJ detection of Baz using antibodies is non-specific.

1.2 Major comments

The manuscript is written in a clear manner, easy to be followed by readers. However, there are some important experimental details that should be provided as the authors advance over previous work regarding Baz localization (points 1.2.1 and 1.2.2). Furthermore, if possible, this reviewer considers that performing the experiment in 1.2.3 would strengthen the authors main message of their manuscript.

1.2.1 Methodology information is missing, and would be necessary to be included for: image acquisition (Objectives, Airyscan mode), image processing (projections, details on linear -e.g. brightness, contrast- or non-linear adjustments of signal -e.g. gamma-). For image processing information, please include it within each figure legend. *

We have added the information regarding objectives and imaging modes to the Materials and Methods section. There it now reads: "Tissues were imaged on a Zeiss LSM880 Airyscan confocal microscope using 25x LCI Plan Neofluar NA 0.8 and 63x Plan Apochromat NA 1.4 oil immersion objectives. If not stated otherwise in the figure legend, all confocal images are single optical sections taken at a pinhole setting of 1 Airy unit. Images were processed with Zen black software (Zeiss) without contrast enhancement. Figures were assembled with Inkscape 1.2 (Inkscape.org) and Powerpoint (Microsoft)."

RNAi experiments lines, temperature for each target and tissue (a table would be helpful) and number of heat shocks performed for FRT/FLP clones.

We have added a table in the Supplementary information giving the precise genotypes for each figure. We have furthermore added the following sentences to the Materials and Methods section: "Crossings for RNAi experiments were set up at 25°C if not indicated otherwise. For generating follicle cell clones in ovaries by Flipase-mediated mitotic recombination of the FRT sites flies were heat shocked for 1h at 37°C 5-7 days prior to preparation of the ovaries. For generation of germ line clones by Flipase-mediated mitotic recombination of the FRT sites flies were heat shocked twice for 2 h at 37°C on two consecutive days in late 2nd, early 3rd instar larval stages."

1.2.2 For each experiment it is unclear the number of specimens (experimental units) and independent experiments that were analysed. It is unclear if the Baz localisation phenotypes are fully penetrant or not as judged by the data provided.

We have added the following section to the Materials and Methods: "Images were analyzed for the presence or absence of a fluorescence signal at the nuclear envelope or the NMJ compared to negative or positive controls, either in the same tissue (mutant clones in the follicular epithelium, RNAi in a specific body wall muscle, junctional versus nuclear signal, anti-Baz staining versus Baz-GFP signal) or in samples processed in parallel (ovaries with follicle cell and germ line clones). Fluorescence intensities were not quantified because the results were obvious and fully penetrant. Therefore, no statistical analysis of the results was required."

1.2.3 This reviewer agrees the data provided strongly suggests the detection of Baz along the nuclear envelope and NMJ is artifactual in the Drosophila tissues that have been studied. However, the nature of the bazEH747 mutant allele is not a deletion of the Baz gene, but instead a nonsense mutation, which, as the authors describe, could potentially generate a small product of 51 aminoacids, corresponding to the N-terminal part of Baz, which is also the target of Baz rabbit antibody ('rb Baz 1-297'). Thus: • Would it be possible to complement the FRT/FLP analyses in the FE using a deficiency that uncovers the baz locus? A persistent detection of Baz signal at the nuclear compartment after complete removal of baz gene products would be an ideal experiment, if feasible.

We agree with the reviewer that the use of a clean deletion allele of the whole baz locus would be the ideal tool for the clonal analysis. However, such an allele does not exist according to our knowledge.

• • Would the authors comment on the possibility the rb Baz antibody 1-297 detect a 51 aminoacids peptide? We consider this possibility very unlikely for two reasons: 1) RNAi affects the baz mRNA and thus should knock down all epitopes to the same degree. However, we see a complete loss of junctional Baz signal but no reduction of the signal at the nuclear envelope or the NMJ upon RNAi targeting baz. 2) The GFP-Baz fusion proteins do not show any signal at the NMJ or the nuclear envelope upon imaging of the native GFP fluorescence or upon antibody staining with an anti GFP antibody, although both the Baz-GFP BAC line and the GFP-Baz protein trap line express full-length Baz including the N-terminal epitope that is potentially still expressed in the bazEH747* allele. We have added a passage summarizing these considerations to the Discussion section.

*1.3 Minor comments

This manuscript is largely based on imaging data. Therefore, it would be beneficial for the ease of comprehension of figure panels:

1.3.1 More general use of insets to show with larger magnification and clarity the data indicated with arrows and arrowheads.*

We have added arrowheads, arrows and additional symbols to point to features of interest in all figure panels where this is helpful.

1.3.2 Using negative grayscale either for insets or single channel data.

We have changed all single channel image panels to negative (inverted) grayscale.

1.3.3 For coloured-overlays please bear in mind using colors that would be suitable for colour-blinded readers.

In all multicolor overlay images, red has been omitted and instead green, magenta, blue and grayscale have been used to improve the visibility for color-blind readers.

1.3.4 Figures showcasing the clonal analyses (both MARCM and FRT/FLP): might be worth indicating the boundaries of clones in single channel data with a dotted line.

We have marked the clone boundaries of the MARCM clones by dashed lines in Fig. 2D, E and have added a high magnification inset to show the clone boundaries (Fig. 2D', E').

Significance

*2 Significance

The findings provided by this manuscript will be of importance for researchers in the field of cell polarity, conducting research on Bazooka/Par-3 and associated proteins, both within the Drosophila field and other model organisms. The present study presents an advance towards a specific and most likely artifactual observation of Par-3/Bazooka. It will help to re-think the tools used for detecting Par-3/Bazooka in different animal models, and in this regard, will be helpful for the community.*

We thank the reviewer for appreciating the importance of this work.

*This work does not focus on Par-3/Bazooka biology, nor provides new insights into Par-3/Bazooka function, however, it is clear for this reviewer the later is not the aim of this manuscript.

Reviewer expertise:

• Drosophila genetics
• Developmental cell biology and morphogenesis
• Cytoskeleton, cell cell adhesion and cell polarity*

Reviewer #3 *(Evidence, reproducibility and clarity (Required)):__

__Kim et al. address a common but frequently neglected problem in molecular and cellular biology: sophisticated tests for the specificity of antibodies. The protein Bazooka (Baz) is a member of the Par complex that usually resides in apicocortical regions of epithelial cells. Several publications, however, report expression in other subcellular compartments or cell types, such as the nuclear lamina or neuromuscular junction (NMJ). The authors have used a panel of polyclonal antibodies, genetic constructs and mutant alleles to show that staining of Baz in the nuclear envelope or NMJ is likely unspecific due to an unknown cross-reactivity. Specifically, four antisera, raised against different GST-Baz fusion proteins in different species, recognized Baz at cortical membranes, around nuclei and at NMJs. Nuclear and NMJ staining, however, persisted in baz-RNAi experiments or baz mutant clones. If the endogenous locus is tagged with GFP, Baz-GFP localized to cortical membranes in imaginal disc epithelial cells but was but not detectable in nuclear envelopes or NMJs in muscles. The authors conclude that they could not find evidence for either nuclear or NMJ localization of Baz and any results derived from these antibodies should be regarded with caution.

The manuscript reports a careful and thorough evaluation of anti-Baz antibodies used in the scientific community. Since it might impact previous findings, any remaining uncertainties should be clarified before publication. I have therefore a number of suggestions to improve the manuscript.

Major comments:

1) Any truncation or addition of amino acids might affect the subcellular localization of proteins. Important molecular information on the baz alleles and GFP-fusion proteins are therefore missing in the manuscript. Specifically, what is the underlying molecular nature of the baz alleles used in the study, e.g. bazEH747 (nonsense? position?)? At which amino acid position and in which protein domain is GFP fused to Baz in Baz-GFP (Bac) and Baz-GFP (Trap)? Would these fusions affect subcellular localization and/or functionality? While the authors positively tested Baz-GFP (Bac) in a baz mutant background, this cannot easily be done for Baz-GFP (Trap). The authors should therefore clarify, e.g. by RT-PCR, which of the four Baz isoforms are fused to GFP in Baz-GFP (Trap) and if this might affect functionality and/or location? This information should be depicted or listed together with the epitopes of the antibodies in a figure or table, respectively, in the main manuscript for better orientation of the reader. *

bazEH747 is a strong loss-of-function allele with a point mutation changing the codon for Q51 to Stop in all four isoforms (numbering is according to isoform A) (Krahn et al., 2010; Shahab et al., 2015). In the Results section, we have changed the wording as follows to make this clear: "For clonal analysis the strong loss-of-function allele bazEH747 was used, where a point mutation in exon 4 results in a premature stop close to the N-terminus of all four isoforms (the codon for amino acid residue Q51 is mutated to a stop in isoform A) (Krahn et al., 2010)."

We have added two additional supplemental figures to precisely show the insertion site of GFP in the GFP-Baz trap line (Fig. S5) and the Baz-GFP BAC line (Fig. S6). We have changed the Results section to precisely explain the nature of the two Baz-GFP lines as follows: "While strong nuclear envelope immunostaining was observed using several independently raised anti Baz antibodies (Fig. 1; Fig. S1), no nuclear envelope localization was detected in follicular epithelial cells and in larval body wall muscles using a Baz-GFP BAC line (Besson et al., 2015) (Fig. S3C-D', S4A, A') nor in a GFP-Baz protein-trap line (Buszczak et al., 2007)(Fig. S3E-F', S4C, C'). In the GFP-Baz protein-trap line an engineered exon encoding for GFP is inserted into the second untranslated exon (Fig. S5). This exon encoding for GFP is predicted to be spliced in frame into the mRNAs RA and RC encoding for isoforms PA and PC whose translation starts in exon 1 (Fig. S5), resulting in insertion of GFP between amino acid residues K40 and P41 of isoforms PA and PC. The transcripts RB and RD encoding Baz isoforms PB and PD have their translation start within exon 3 and thus cannot form fusion proteins with GFP inserted in exon 2 (Fig. S5). However, GFP-Baz protein trap flies are homozygous viable and are phenotypically indistinguishable from wild type flies, indicating that the corresponding GFP fusion protein is fully functional and faithfully reflects the expression pattern and subcellular localization of Baz isoforms PA and PC. The BAC line integrates the GFP within exon 10 between amino acid residues L1424 and Q1425 of isoform PA, giving rise to GFP fusion proteins for all four isoforms (Fig. S6) (Besson et al., 2015). Like the protein-trap GFP-Baz fusion protein, the Baz-GFP fusion protein in the BAC line is fully functional as it completely rescued lethality and fertility of the bazEH747 allele (Fig. S7D-D') and the baz815-8 allele (Besson et al., 2015)."

*2) Figure 3D-G: The images for Baz-GFP nicely show that GFP is expressed in imaginal discs but not at NMJs. However, when brightness of Fig. 3D' and 3F' is increased nuclear envelopes, tracheal branches and some synaptic boutons are clearly visible in the Baz-GFP channels. These are likely background signals due to the staining procedure, but to avoid any confusion, images showing unstained (native) GFP fluorescence should be included to proof that there are no residual signals. GFP fluorescence survives formaldehyde fixation and many GFP exon traps are clearly visible even in the absence of immunofluorescent stainings. Furthermore, Fig. 3G appears vastly different compared to Fig. 3E and Baz localization at cell-cell junctions cannot be recognized by people unfamiliar with imaginal discs. The images in Fig. 3G are therefore not suitable and should be replaced. *

We have added the new Fig. S4 showing the GFP signal without antibody staining of somatic body wall muscles and wing imaginal discs of larvae expressing the Baz-GFP BAC and GFP-Baz trap transgenes. We have also replaced Fig. 3G with images that can easily be compared with the images in Fig. 3E. The following paragraph was added to the Results section: "These findings were confirmed by analysis of fixed larval tissues that were imaged for GFP fluorescence without anti GFP antibody staining (Fig. S4). Neither in the Baz-GFP BAC line (Fig. S4A, A'), nor in the GFP-Baz trap line (Fig. S4C, C') any nuclear envelope or NMJ signal was detectable in somatic muscles, whereas junctional signal in wing imaginal discs was readily detectable in both lines (Fig. S4B, D)."

*3) The argument that baz4 and baz815-8 carry second site mutations is not fully convincing (page 10, 13). Why should two independent baz alleles carry an additional hit that affect Spectrin levels? Other explanations might be possible. While downregulation of Baz in muscles by RNAi is a good approach to tackle the question of Spectrin localization and expression levels, RNAi itself has its own uncertainties. Why not showing the effect on Spectrin levels or the lack of Baz at the NMJ (or the nuclear envelopes) in "clean" baz null embryos or larvae (e.g. bazEH747/Df)? NMJs can be stained in late stage embryos or compound heterozygous null mutants quite frequently survive until larval stages. *

We do not have a good explanation for the published reduction of Baz and a-Spectrin signal at the NMJ in larvae heterozygous for the baz alleles baz4 and baz815-8 (Ruiz-Canada et al., 2004; Ramachandran et al., 2009), as our analysis shows that Baz is not expressed there, rendering the reported phenotypes very difficult to explain. It is beyond the scope of our paper to proof that the data published by Ruiz-Canada et al. (2004) and Ramachandran et al. (2009) are indeed reproducible. Our speculation that second site hits on these two mutant chromosomes may have caused the published effects is just based on our own published observation that commonly used chromosomes with these two mutant baz alleles have stronger phenotypes than a clean baz loss-of-function allele (Shahab et al., 2015). We have changed the wording of the corresponding paragraph as follows: "It has been published that heterozygous baz4 mutant larvae show a significant decrease in immunofluorescence signal of Baz and also of Spectrin at the NMJ (Ruiz-Canada et al., 2004). Another publication showed a significant decrease in Baz and Spectrin immunostaining at the NMJ of larvae heterozygous for the baz815-8 allele (Ramachandran et al., 2009). We did not attempt to reproduce these findings. However, in our hands mitotic clones generated with FRT chromosomes carrying these latter two baz alleles showed polarity phenotypes in the follicular epithelium, whereas clones of the clean bazEH747 null allele did not show any polarity defect (Shahab et al., 2015), raising the possibility that the NMJ phenotypes observed by Ruiz-Canada et al. (2004) and Ramachandran et al. (2009) were caused by second site mutations on these chromosomes rather than by reduced Baz activity.

bazEH747 hemizygous mutant embryos are so abnormal and malformed at late embryonic stages that we did not attempt to stain these for Baz immunoreactivity at NMJs.

4) It is not really made clear in the manuscript, why the additional reactivity of the anti-Baz antibodies has not been noticed earlier. The paper should therefore include a summarizing paragraph that describes how the specificities of the antibodies have been tested in the past in the laboratories that used them. Have they never been tested in null mutant animals? In null mutants it should be obvious to determine, if some staining patterns do not disappear.

The vast majority of publications on Baz including those from our own laboratory focused on the functions of Baz at junctions and in the control of cell polarity. For these functions the cortical localization of Baz is relevant, which has been shown to be specific in many independent studies using null alleles and RNAi. Only few publications, in particular those from the laboratory of Vivian Budnik, have focused on potential functions of Baz at the NMJ and the nuclear envelope. Why in these studies no convincing proof of the specificity of the signal at those "unconventional" locations has been provided is beyond our knowledge.

5) Figure 4 is very difficult to comprehend and should be better labeled (e.g. anterior-posterior, dorsal-ventral, muscle fibers, unspecific signals). It is standard in the field to show ventral muscles 12, 13 or 6, 7 in the center of the image and in a similar orientation (anterior left, dorsal up). Better images should be shown.

We understand that for researchers interested in the function of specific muscles it is important to adhere to conventions regarding the orientation of muscles in figures. However, in our case it is just relevant whether a muscle expresses RNAi against a gene of interest (GFP+) or not (GFP-) in order to compare the signal intensity for Baz and Spectrin in these two situations. Thus, although we appreciate the validity of this comment, we decided to leave the original images unchanged. However, to help the reader in identifying relevant structures more easily, we have added color-coded arrows and arrowheads to mark NMJs and nuclear envelopes in GFP+ and GFP- muscles.

*Reviewer #3 (Significance (Required)):

The authors provide a critical assessment on the specificity of antibodies and highlight the necessity to carefully test antibodies and the conclusions drawn from the resulting stainings, especially when antibodies are bought from companies or have previously been published as specific. This is extremely important for the interpretation of experiments in all fields of molecular and cellular biology. *

We thank the reviewer for appreciating the importance of this work.

Note: This preprint has been reviewed by subject experts for Review Commons. Content has not been altered except for formatting.

Learn more at Review Commons

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Referee #3

Evidence, reproducibility and clarity

Kim et al. address a common but frequently neglected problem in molecular and cellular biology: sophisticated tests for the specificity of antibodies. The protein Bazooka (Baz) is a member of the Par complex that usually resides in apicocortical regions of epithelial cells. Several publications, however, report expression in other subcellular compartments or cell types, such as the nuclear lamina or neuromuscular junction (NMJ). The authors have used a panel of polyclonal antibodies, genetic constructs and mutant alleles to show that staining of Baz in the nuclear envelope or NMJ is likely unspecific due to an unknown cross-reactivity. Specifically, four antisera, raised against different GST-Baz fusion proteins in different species, recognized Baz at cortical membranes, around nuclei and at NMJs. Nuclear and NMJ staining, however, persisted in baz-RNAi experiments or baz mutant clones. If the endogenous locus is tagged with GFP, Baz-GFP localized to cortical membranes in imaginal disc epithelial cells but was but not detectable in nuclear envelopes or NMJs in muscles. The authors conclude that they could not find evidence for either nuclear or NMJ localization of Baz and any results derived from these antibodies should be regarded with caution.

The manuscript reports a careful and thorough evaluation of anti-Baz antibodies used in the scientific community. Since it might impact previous findings, any remaining uncertainties should be clarified before publication. I have therefore a number of suggestions to improve the manuscript.

Major comments:

1. Any truncation or addition of amino acids might affect the subcellular localization of proteins. Important molecular information on the baz alleles and GFP-fusion proteins are therefore missing in the manuscript. Specifically, what is the underlying molecular nature of the baz alleles used in the study, e.g. bazEH747 (nonsense? position?)? At which amino acid position and in which protein domain is GFP fused to Baz in Baz-GFP (Bac) and Baz-GFP (Trap)? Would these fusions affect subcellular localization and/or functionality? While the authors positively tested Baz-GFP (Bac) in a baz mutant background, this cannot easily be done for Baz-GFP (Trap). The authors should therefore clarify, e.g. by RT-PCR, which of the four Baz isoforms are fused to GFP in Baz-GFP (Trap) and if this might affect functionality and/or location? This information should be depicted or listed together with the epitopes of the antibodies in a figure or table, respectively, in the main manuscript for better orientation of the reader.
2. Figure 3D-G: The images for Baz-GFP nicely show that GFP is expressed in imaginal discs but not at NMJs. However, when brightness of Fig. 3D' and 3F' is increased nuclear envelopes, tracheal branches and some synaptic boutons are clearly visible in the Baz-GFP channels. These are likely background signals due to the staining procedure, but to avoid any confusion, images showing unstained (native) GFP fluorescence should be included to proof that there are no residual signals. GFP fluorescence survives formaldehyde fixation and many GFP exon traps are clearly visible even in the absence of immunofluorescent stainings. Furthermore, Fig. 3G appears vastly different compared to Fig. 3E and Baz localization at cell-cell junctions cannot be recognized by people unfamiliar with imaginal discs. The images in Fig. 3G are therefore not suitable and should be replaced.
3. The argument that baz4 and baz815-8 carry second site mutations is not fully convincing (page 10, 13). Why should two independent baz alleles carry an additional hit that affect Spectrin levels? Other explanations might be possible. While downregulation of Baz in muscles by RNAi is a good approach to tackle the question of Spectrin localization and expression levels, RNAi itself has its own uncertainties. Why not showing the effect on Spectrin levels or the lack of Baz at the NMJ (or the nuclear envelopes) in "clean" baz null embryos or larvae (e.g. bazEH747/Df)? NMJs can be stained in late stage embryos or compound heterozygous null mutants quite frequently survive until larval stages.
4. It is not really made clear in the manuscript, why the additional reactivity of the anti-Baz antibodies has not been noticed earlier. The paper should therefore include a summarizing paragraph that describes how the specificities of the antibodies have been tested in the past in the laboratories that used them. Have they never been tested in null mutant animals? In null mutants it should be obvious to determine, if some staining patterns do not disappear.
5. Figure 4 is very difficult to comprehend and should be better labeled (e.g. anterior-posterior, dorsal-ventral, muscle fibers, unspecific signals). It is standard in the field to show ventral muscles 12, 13 or 6, 7 in the center of the image and in a similar orientation (anterior left, dorsal up). Better images should be shown.

Significance

The authors provide a critical assessment on the specificity of antibodies and highlight the necessity to carefully test antibodies and the conclusions drawn from the resulting stainings, especially when antibodies are bought from companies or have previously been published as specific. This is extremely important for the interpretation of experiments in all fields of molecular and cellular biology.

Note: This preprint has been reviewed by subject experts for Review Commons. Content has not been altered except for formatting.

Learn more at Review Commons

---

Referee #2

Evidence, reproducibility and clarity

1. Evidence, reproducibility and clarity

1.1 Summary

This reviewer acknowledges the expertise and contributions of Prof. Wodarz and his research group in the field of development, cell polarity regulation and Drosophila genetics.

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Manuscript summary:

Kim S. et al. explored the localisation of Bazooka, the Drosophila homolog of the polarity protein Par-3, at two non-canonical positions for a cell polarity factor: the nuclear envelope in epithelial tissues and the postsynaptic membrane of the neuromuscular junction (NMJ). Previous work has shown the detection of Par-3/Baz at the nuclear envelope and the NMJ using antibodies against Par-3/Baz. Here, the authors used a combination of genetic perturbations (baz RNAi and generation of genetic mosaics for baz) and GFP-labelled Bazooka lines to test if the antibody-mediated detection of Baz at the nuclear envelope and NMJ is artifactual. The data provided by the authors strongly suggest both the nuclear envelope and NMJ detection of Baz using antibodies is non-specific.

1.2 Major comments

The manuscript is written in a clear manner, easy to be followed by readers. However, there are some important experimental details that should be provided as the authors advance over previous work regarding Baz localization (points 1.2.1 and 1.2.2). Furthermore, if possible, this reviewer considers that performing the experiment in 1.2.3 would strengthen the authors main message of their manuscript.

1.2.1 Methodology information is missing, and would be necessary to be included for: image acquisition (Objectives, Airyscan mode), image processing (projections, details on linear -e.g. brightness, contrast- or non-linear adjustments of signal -e.g. gamma-). For image processing information, please include it within each figure legend. RNAi experiments lines, temperature for each target and tissue (a table would be helpful) and number of heat shocks performed for FRT/FLP clones.

1.2.2 For each experiment it is unclear the number of specimens (experimental units) and independent experiments that were analysed. It is unclear if the Baz localisation phenotypes are fully penetrant or not as judged by the data provided.

1.2.3 This reviewer agrees the data provided strongly suggests the detection of Baz along the nuclear envelope and NMJ is artifactual in the Drosophila tissues that have been studied. However, the nature of the bazEH747 mutant allele is not a deletion of the Baz gene, but instead a nonsense mutation, which, as the authors describe, could potentially generate a small product of 51 aminoacids, corresponding to the N-terminal part of Baz, which is also the target of Baz rabbit antibody ('rb Baz 1-297').

Thus: - Would it be possible to complement the FRT/FLP analyses in the FE using a deficiency that uncovers the baz locus? A persistent detection of Baz signal at the nuclear compartment after complete removal of baz gene products would be an ideal experiment, if feasible. - Would the authors comment on the possibility the rb Baz antibody 1-297 detect a 51 aminoacids peptide?

1.3 Minor comments

This manuscript is largely based on imaging data. Therefore, it would be beneficial for the ease of comprehension of figure panels:

1.3.1 More general use of insets to show with larger magnification and clarity the data indicated with arrows and arrowheads.

1.3.2 Using negative grayscale either for insets or single channel data.

1.3.3 For coloured-overlays please bear in mind using colors that would be suitable for colour-blinded readers.

1.3.4 Figures showcasing the clonal analyses (both MARCM and FRT/FLP): might be worth indicating the boundaries of clones in single channel data with a dotted line.

Referees cross-commenting

I consider that all points/questions raised by other reviewers are fair, in some cases complement this reviewer's points, and in some others coincide. I recommend that all points raised by reviewers #1 and #3 are fully addressed by the authors.

Significance

The findings provided by this manuscript will be of importance for researchers in the field of cell polarity, conducting research on Bazooka/Par-3 and associated proteins, both within the Drosophila field and other model organisms.

The present study presents an advance towards a specific and most likely artifactual observation of Par-3/Bazooka. It will help to re-think the tools used for detecting Par-3/Bazooka in different animal models, and in this regard, will be helpful for the community.

This work does not focus on Par-3/Bazooka biology, nor provides new insights into Par-3/Bazooka function, however, it is clear for this reviewer the later is not the aim of this manuscript.

Reviewer expertise:

• Drosophila genetics
• Developmental cell biology and morphogenesis
• Cytoskeleton, cell cell adhesion and cell polarity

Note: This preprint has been reviewed by subject experts for Review Commons. Content has not been altered except for formatting.

Learn more at Review Commons

---

Referee #1

Evidence, reproducibility and clarity

Baz/Par3 is an important conserved protein acting as a master regulator of cell polarity in a wide range of cell types. This study focuses on re-assessing the subcellular localisation of Baz/Par3 in a range of Drosophila tissues. This is an important study with respect to our understanding of Baz/Par3, as there have been conflicting reports on the localisation of Par complex members - while the majority show localisation to cell cortex and intercellular junctions, several reports have claimed that Par complex members localise at additional subcellular sites including the nucleus, nuclear envelope and neuromuscular junction. In this study the authors re-assess this issue for Baz/Par3 in a comprehensive and thorough manner.

1. They used a variety of antibodies raised in different host animals against different epitopes of Baz
2. They tested the specificity of these antisera using mosaic analysis with null mutant baz alleles and tissue-specific RNAi against baz
3. They used a GFP-tagged Baz under control of its endogenous promoter in a baz null mutant background to compare with the subcellular localisation of the respective GFP-Baz fusion proteins to the staining results with anti-Baz antisera

The data from each of these experiments are very clear and convincing. Comprehensive methods are included which means that each of the experiments with specific anti-sera/RNAi lines/GFP-tagged conditions could be reproduced. There are a couple of experiments which were performed in support of the conclusions (extra RNAi lines and stronger expression of Gal4) listed as (data not shown). I would strongly suggest including these data as extra supplemental figures. Together, their results clearly show that Baz/Par3 localises to the cortex and intercellular junctions, but that anti-sera staining at the NMJs and nuclear envelope appear to be a staining artifact, likely due to staining with an unidentified epitope.

Minor comments

1. Many of the figures have overlays of red and green which will be indistinguishable from each other to colour-blind readers. Please alter to make colour-blind friendly (eg magenta-green)
2. In Fig 2D please indicate where the epidermis and neuroblasts are
3. In the following two places there are experiments describe where the data is listed as not shown. Please show the data as additional supplemental data. They are P8 - This result was confirmed using the CY2::Gal4 driver line expressed in the follicular epithelium and with three different RNAi lines against baz (data not shown). P11 - We also did not see any downregulation of Baz or aspectrin upon baz-RNAi in M12 at 29{degree sign}C, when the UAS-Gal4 system is maximally active (data not shown).
4. Figure 3 - this would be easier to interpret with a few arrows/arrowheads indicating the NMJs

Significance

It will be important to publish these results as it means that findings for a function of Baz/Par3 at the NJM and the nuclear envelope should be regarded with caution, and it may save researchers chasing for functions for Baz/Par3 in places where they are simply not expressed. As much of our fundamental understanding of how Par3 works in vertebrates has its roots in studies in Drosophila, this is likely to be of wide relevance.

Point of view- I think this sections showcases the point of view of RFK Jr winning the election because it talks about his limited abilities to get on the ballots across all states while also comparing RFK Jr's chances of winning to Biden or Trump.

Purpose- I think this section shows the purpose of this article which is to figure out if RFK Jr, is capable of winning the election and the article further explain who RFK Jr is and if he is a capable elective to be voted for.

A Useful Productivity Measure? by [[James Shore]]

This passage discusses two tactics for fostering a conducive environment for mentorship or collaboration within a professional setting. Firstly, it emphasizes the importance of approachability by adopting an "open-door" policy, enabling colleagues to initiate conversations spontaneously for discussing issues, projects, or seeking guidance. Secondly, it underscores the significance of embracing feedback as a crucial component of personal and professional development, despite the potential emotional discomfort it may evoke. Encouraging individuals to reflect on the purpose behind feedback can aid in fostering a constructive mindset conducive to learning and growth.

é uma modalidade de extinção unilateral dos contratos de concessão de obras públicas e serviços públicos.

Do reaching sustainable development goals and solving global challenges need all of these models?

What do we need to understand robustness of function and biodiversity and services? Is the best topology simply one that can plug into something dynamic?

Do you think the ADBM is close to this?

+ non-trophic interactions.

see comment above - I think we are realising we kind of need a list of 'things people do with food webs'....

this feels like a criticism rather than a fact, and I think it's more of a fact?

Also, are we declaring TG and IP 'classes' of models

is this an example of the distinction between TG and IP?

I've lost the reference here... which? TG or IP?

available, necessary and sufficient?

see above comment...

I like this. I wonder if we can link it more closely to biodiversity per se? And that question of what are we doing with a network... perhaps too something about the value of a network.... biodiversity becomes a structured value, connectivity is enumerated and tied to stability, indirect effects (motifs!) become apparent/important, secondary extinctions become measurable, community/biodiversity responses become measurable.

does this mass effect equate with the first principle of probability? In foraging biology, the default for preference is in proportion to abundance, and I think this is kind of what this means for edge weight? Not sure it's how, but how much?

Super, but perhaps more effective in reverse?

The ingredients one thus uses to construct networks of nodes and edges generates unique representations of mechanism (recipes) that allow inference and reasoning about the structure, aspects of dynamics (e.g. stability) and potentially the function of communities (e.g. flux).

would it be better to phrase as an opportunity to do more cleverer thingies?

if possible, we should state it, rather than refer to it... if it's a simple argument for ....

shall we say strength of interaction?

not sure links are overcorrected.... the link density is not representative of something?

is this quantify, or specify? One some of those are quantification, other are just existence?

Of course the species node IS an aggregation in and of itself, often averaging over life stages/ages (e.g. 10.1002/ecy.2523)

I am still getting my head around this.... does this include weights as interaction strengths? Is that probabilistic?

would it be among as there can be links from A to many?

Veilleux, Jennifer C. 2013. “The Human Security Dimensions of Dam Development: The Grand Ethiopian Renaissance Dam.” Global Dialogue 15 (2): 41–52.

I try to do this in as human a way as possible at the beginning of a semester and establish boundaries in my 'communication policy'. I tell students they are welcome to contact me during whatever hours suits them, but to know that I will respond in the regular working day as I have young children at home who need me to be present

I love this idea of meeting the student where they are at, Mastery Pathways on Canvas is a great option to explore this

From a college perspective, I have some questions on this: how do we manage this within an 8 week or 16 week block of classes? How does this work for students with AD(H)D or delayed executive functioning? Research shows they need deadlines and grade penalties for late assignments are currently recommended to prevent procrastination, several missed assignments and student overwhelm

Abdelhady, Dalia, Karin Aggestam, Dan-Erik Andersson, Olof Beckman, Ronny Berndtsson, Karin Broberg Palmgren, Kaveh Madani, Umut Ozkirimli, Kenneth M. Persson, and Petter Pilesjö. 2015. “The Nile and the Grand Ethiopian Renaissance Dam: Is There a Meeting Point between Nationalism and Hydrosolidarity?” Journal of Contemporary Water Research & Education 155 (1): 73–82. doi:10.1111/j.1936-704X.2015.03197.x. , 73

Scott, James 1999. Seeing Like a State: How Certain Schemes to Improve the Human Condition Have Failed. Yale University Press.

wow i didnt know there was private features on schoology!

The Education Department initially placed T.J. in a middle school in East Flatbush, but his lawyer, Mr. Hochbaum, had concerns. He felt that T.J. would be the only student with an intellectual disability in his class and that the school's curriculum wouldn't meet his needs. The school is also part of a program for struggling schools. In the 2017-18 school year, only 13 percent of students passed the state math test, and 28 percent were proficient in reading. This highlights the challenges T.J. and his family are facing in finding a school that is the right fit for him.

This passage discusses New York City's Early Intervention program for children up to age 3 and the Committee on Preschool Special Education for children aged 3 to 5. In T.J.'s case, there seems to have been a gap in services. Despite being evaluated and recommended for therapy and special education services before age 3, it's unclear if he ever received them. His mother, Kerrin, stated that he received no services after turning 3. This highlights the challenges families face in navigating the system to ensure their children receive needed support. The mention of the family's lawyer struggling to obtain records adds to the bureaucratic hurdles involved.

This passage shares the story of T.J., a 12-year-old boy in New York City's special education system. His journey began when his mother, Kerrin, sought help for him due to learning challenges. Despite his friendly nature and good manners, T.J. struggles with reading and writing, still performing at a first-grade level in sixth grade. This has left him feeling left behind as his classmates move ahead. The passage highlights the challenges faced by T.J. and his family in navigating the educational system, showing the impact it has on T.J.'s education and well-being.

I believe this is meant to evoke an emotional response in the reader since, while it is true, there is no hyper-link to an instance of this occurring or a citation to fact check so in order to verify this information you have to do your own research outside the article. Without citation this information has to be processed as a persuasive opinion relying on a gut response to convince readers to agree with it.

This passage discusses the issue of students of color being disproportionately placed in special education. It notes that while this is a complex and controversial issue, it's important to ensure that all students who need special education services receive them. The passage suggests that biased placement and segregation can be reduced by implementing culturally responsive practices in schools. This includes providing training to increase cultural competence among educators and ensuring that decision-making committees are diverse. Additionally, the passage emphasizes the need to adequately fund schools in low-income areas to provide support for all students.