4,197 Matching Annotations
  1. Aug 2021
    1. found that NLRP3 overexpression inhibits cell proliferation and stimulates apoptosis in leukemic cells.
    2. NLRP3 enhances IL-1beta , subsequently activating NF-kappaB , and initiates JNK signaling to cause proliferation and invasion in gastric cancer ( 21 ) .

      NLRP3 activates NFkappaB.

    3. NLRP3 in the primary lesion of cancer cells drives the production of pro-IL-1beta, DC maturation, and the secretion of IL-1beta to support the evolution of tumor specific CD8 + T cells.

      NLRP3 activates CD8.

    4. In the primary lesion of cancer cells, NLRP3 drives the production of pro-IL-1beta, DC maturation, and the secretion of IL-1beta to support the differentiation of tumor specific CD8 + T cells.

      NLRP3 activates CD8.

    5. Moreover , NLRP3 downstream , IL-1beta , also stimulates the production of ROS that , in turn , induces DNA damage and cancer development in CRC ( 42 ) ( Table 2 ) .
    6. NLRP3 inflammasomes mediate both suppressions of apoptosis and progression of the cell cycle by leptin dependent ROS production in breast cancer, which is mediated via estrogen receptor alpha (ERalpha)/reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase signaling.
    7. Moreover, NLRP3 downstream, IL-1beta, also stimulates the production of ROS that, in turn, induces DNA damage and cancer development in CRC (XREF_TABLE).
    8. Although caspase-1 activation is the major downstream event of NLRP3 inflammasome assembly, recent studies have reported that NLRP3 inflammasome could also be activated by caspase-8.

      CASP8 activates NLRP3.

    9. ATP, one of the major cancer metabolites and constituents of the TME, serves as a key DAMP that activates NLRP3 inflammasome via the purinergic P2X7 receptors.
    10. Among the known human chemokines, a co-regulated set of four (chemokine (C-C motif) ligand (CCL)-4, CCL-5, chemokine (C-X-C motif) ligand (CXCL)-9, CXCL-10) chemokines is upregulated in primary PDA carcinoma and PDA liver metastasis, which regulates CD8 + T cell infiltration, activates T cells, and promotes NLRP3 mediated T cell priming and enhances anti-tumor CD8 + T cell cytotoxic activity for an effective immune checkpoint therapy response.
    1. The probability for the occurrence of sulphur plumes is enhanced in years with a lower annual mean of upwelling intensity, decreased oxygen supply associated with decreased lateral ventilation of bottom waters, more southern position of the Angola Benguela Frontal Zone, increased mass fraction of South Atlantic Central Water and stronger downwelling coastal trapped waves.

      dioxygen activates water.

    1. Interestingly, in addition to its role in inhibiting caspase-1, parthenolide also directly inhibits NLRP3 by inhibiting its ATPase activity, which is required for activation of caspase-1.

      parthenolide inhibits NLRP3.

    2. XREF_BIBR It was thought to be an NFkappaB inhibitor through selective inhibition of kappaB kinase beta (IKKbeta) kinase activity; however, in addition to its effects on NFkappaB activation, parthenolide has now been shown to inhibit activation of caspase-1 in response to NLRP3, NLRP1, and NLRC4 stimulation.

      parthenolide inhibits CASP1.

    3. The effect of parthenolide on the spectrum of inflammasomes implies that it acts on a common component and, accordingly, it has been shown that parthenolide is a direct inhibitor of caspase-1, causing alkylation of caspase-1 on a number of cysteine residues.

      parthenolide inhibits CASP1.

    1. IL-1beta secretion was not affected by treatment with the NLRP3 inhibitor glyburide XREF_BIBR or parthenolide, which has also been shown to inhibit NLRP3 XREF_BIBR.

      parthenolide inhibits NLRP3.

    2. Glyburide and parthenolide both inhibited NLRP3 activation by LPS and ATP (data not shown).

      parthenolide inhibits NLRP3.

    3. XREF_BIBR have shown that parthenolide directly inhibits caspase-1 by alkylation of certain cysteine residues.

      parthenolide inhibits CASP1.

    4. However, the previous study xref did not test AIM2 activation so perhaps parthenolide only inhibits caspase-1 in response to NLRP3 or NLRC4 activation.

      parthenolide inhibits CASP1.

    5. Our results disagree with this assertion as we did not find that parthenolide inhibited caspase-1 in response to AIM2 stimulation.

      parthenolide inhibits CASP1.

    1. Water fluxes through pressurized root systems treated with nitrogen and low oxygen (< 2% O (2)), elevated CO (2) (20% CO (2)), and low O (2) with elevated CO (2) concentrations were reduced to 40, 51 and 58%, respectively, of J (v) of plants aerated with ambient air.

      dioxygen activates water.

    1. Knockout of TGF-β receptor 2 in hair follicle melanocyte lineage blocks the Smad2 phosphorylation, resulting in a loss of quiescence state of McSCs.

      SMAD2 is phosphorylated.

    2. TGF-β binds TGF-β receptors in melanocytes, leading to the phosphorylation of downstream effector Smad2, which inhibits melanocyte growth and melanogenesis through downregulating PAX3 and MITF transcription xref , xref .

      SMAD2 is phosphorylated.

    3. On the contrary, when agouti activity is inhibited by beta-Defensin, or Corin, or beta-catenin in the DP, the yellow coat turns to be black XREF_BIBR - XREF_BIBR.

      beta-Defensins inhibits agouti.

    4. Conditional knockout of NFIB in HFSCs promotes McSCs proliferation and differentiation, indicating a role of NFIB as a regulator of McSC behavior 73.
    5. Conditional knockout of NFIB in HFSCs promotes McSCs proliferation and differentiation, indicating a role of NFIB as a regulator of McSC behavior 73.
    6. On the contrary, when agouti activity is inhibited by beta-Defensin, or Corin, or beta-catenin in the DP, the yellow coat turns to be black XREF_BIBR - XREF_BIBR.

      DSP inhibits agouti.

    7. On the contrary, when agouti activity is inhibited by beta-Defensin, or Corin, or beta-catenin in the DP, the yellow coat turns to be black XREF_BIBR - XREF_BIBR.

      CTNNB1 inhibits agouti.

    8. On the contrary, when agouti activity is inhibited by beta-Defensin, or Corin, or beta-catenin in the DP, the yellow coat turns to be black XREF_BIBR - XREF_BIBR.

      CORIN inhibits agouti.

    9. Mechanistically, knockout of Kindlin-1 promotes cutaneous epithelial stem cells differentiation via inhibiting alpha (v) beta (6) integrin mediated TGF-beta1 liberation and promoting integrin independent Wnt ligand expression to activate Wnt and beta-catenin signaling 82.

      TGFB1 inhibits Integrins.

    10. Mechanistically, knockout of Kindlin-1 promotes cutaneous epithelial stem cells differentiation via inhibiting alpha (v) beta (6) integrin mediated TGF-beta1 liberation and promoting integrin independent Wnt ligand expression to activate Wnt and beta-catenin signaling 82.

      Wnt inhibits CTNNB1.

    11. Mechanistically, knockout of Kindlin-1 promotes cutaneous epithelial stem cells differentiation via inhibiting alpha (v) beta (6) integrin mediated TGF-beta1 liberation and promoting integrin independent Wnt ligand expression to activate Wnt and beta-catenin signaling 82.

      Wnt inhibits Wnt.

    12. Inhibition of Wnt signaling by a Wnt antagonist secreted frizzled-related protein 4 (sFRP4), which is exclusively expressed in the epithelial cells but not the melanocytes of the hair follicle, results in a decrease of melanocytes differentiation in the regenerating hair follicle xref .

      antagonist inhibits signaling.

    13. Even the melanocyte progenitors emigrate into feathers, the differentiation may be suppressed by agouti, made by the peripheral pulp fibroblasts 1.

      Agouti inhibits cell differentiation.

    14. Pax3 not only promotes melanogenesis by activating the expression of MITF, but also maintains McSCs quiescence by competing with MITF through binding an enhancer responsible for the expression of dopachrome tautomerase (DCT), an intermediate in the biosynthesis of melanin.

      PAX3 increases the amount of MITF.

    15. Alx3 decreases melanin production by directly suppressing the expression of MITF, by indirectly inhibiting the secretion of Edn3, and by indirectly promoting the expression of ASIP.

      ALX3 increases the amount of MITF.

    16. Sox10, Pax3, and Wnt3a mediated Wnt and beta-catenin signaling induce the transcription of MITF and promote differentiation of neural crest into melanoblasts XREF_BIBR - XREF_BIBR, though MITF is not expressed in the neural crest.

      WNT3A increases the amount of MITF.

    17. Inhibition of NFIB signaling in HFSCs directly stimulates expression of endothelin 2 (Edn2), which is required in HFSCs dependent McSCs activation.

      NFIB decreases the amount of EDN2.

    18. For example, MITF dependent expression of microRNA-211 promotes pigmentation in melanoblast and melanocyte cell lines by inhibiting the expression of TGF-beta receptor 2, which is involved in the maintenance of McSCs quiescence.

      Modified MITF decreases the amount of TGFBR.

    19. Alx3 decreases melanin production by directly suppressing the expression of MITF, by indirectly inhibiting the secretion of Edn3, and by indirectly promoting the expression of ASIP.

      ALX3 decreases the amount of ASIP.

    20. For example, MITF dependent expression of microRNA-211 promotes pigmentation in melanoblast and melanocyte cell lines by inhibiting the expression of TGF-beta receptor 2, which is involved in the maintenance of McSCs quiescence.

      Modified MIR211 decreases the amount of TGFB.

    21. For example, MITF dependent expression of microRNA-211 promotes pigmentation in melanoblast and melanocyte cell lines by inhibiting the expression of TGF-beta receptor 2, which is involved in the maintenance of McSCs quiescence.

      Modified MIR211 decreases the amount of TGFBR.

    22. Through interacting with PAX3, FOXD3 prevents binding of PAX3 to MITF promoter to repress melanogenesis in zebrafish, quail and chick neural crest cells XREF_BIBR, XREF_BIBR, suggesting that down-regulation of Foxd3 is a crucial step during the early phase of melanoblast lineage specification from neural crest cells.

      MITF binds PAX3.

    23. Through interacting with PAX3, FOXD3 prevents binding of PAX3 to MITF promoter to repress melanogenesis in zebrafish, quail and chick neural crest cells xref , xref , suggesting that down-regulation of Foxd3 is a crucial step during the early phase of melanoblast lineage specification from neural crest cells.

      MITF binds PAX3.

    24. In human melanoma cells, MITF also interacts directly with beta-catenin and redirects beta-catenin transcriptional activity away from target genes regulated by Wnt and beta-catenin signaling, toward MITF specific target promoters to activate transcription 57.

      CTNNB1 binds MITF.

    25. In human melanoma cells, MITF also interacts directly with β-catenin and redirects β-catenin transcriptional activity away from target genes regulated by Wnt/β-catenin signaling, toward MITF-specific target promoters to activate transcription xref .

      CTNNB1 binds MITF.

    26. TGF-β binds TGF-β receptors in melanocytes, leading to the phosphorylation of downstream effector Smad2, which inhibits melanocyte growth and melanogenesis through downregulating PAX3 and MITF transcription xref , xref .

      TGFB binds SMAD2.

    27. TGF-beta binds TGF-beta receptors in melanocytes, leading to the phosphorylation of downstream effector Smad2, which inhibits melanocyte growth and melanogenesis through downregulating PAX3 and MITF transcription XREF_BIBR, XREF_BIBR.

      TGFB binds TGFBR.

    28. Notch ligands including Jagged1, Jagged2, Delta-like1, Delta-like3 and Delta-like4 bind to Notch receptor, which induces signal transduction cascade through the induction of transcription factor RBP-JK to initiate the transcription of target genes 69.

      Notch binds Notch.

    29. UV and ionizing radiation induced DNA damage triggers McSC differentiation, leading to McSC exhaustion and hair graying XREF_BIBR, XREF_BIBR, XREF_BIBR.
    30. Although alpha-MSH can derive from epithelial cells and systematically, the activation of Mc1r signaling by alpha-MSH is in the DP, which further regulates melanocyte pigmentation.

      POMC activates MC1R.

    31. These data indicate that MITF may enhance the role of Wnt and beta-catenin signaling in proliferation and differentiation of McSCs in a feedback mechanism.

      MITF activates CTNNB1.

    32. Sox10, Pax3, and Wnt3a mediated Wnt and beta-catenin signaling induce the transcription of MITF and promote differentiation of neural crest into melanoblasts XREF_BIBR - XREF_BIBR, though MITF is not expressed in the neural crest.
    33. These data indicate that MITF may enhance the role of Wnt and beta-catenin signaling in proliferation and differentiation of McSCs in a feedback mechanism.

      MITF activates Wnt.

    34. HGF, SCF, and End3 have been revealed to promote melanoblast or melanocyte proliferation and differentiation XREF_BIBR, XREF_BIBR.
    35. HGF, SCF, and End3 have been revealed to promote melanoblast or melanocyte proliferation and differentiation XREF_BIBR, XREF_BIBR.
    36. HGF, SCF, and End3 have been revealed to promote melanoblast or melanocyte proliferation and differentiation XREF_BIBR, XREF_BIBR.
    37. HGF, SCF, and End3 have been revealed to promote melanoblast or melanocyte proliferation and differentiation XREF_BIBR, XREF_BIBR.
    38. Continuous activation of beta-catenin Signaling in McSCs promotes McSCs differentiation, exhaustion and premature hair graying 8.
    39. In human melanoma cells, MITF also interacts directly with beta-catenin and redirects beta-catenin transcriptional activity away from target genes regulated by Wnt and beta-catenin signaling, toward MITF specific target promoters to activate transcription 57.
    40. Sox10, Pax3, and Wnt3a mediated Wnt and beta-catenin signaling induce the transcription of MITF and promote differentiation of neural crest into melanoblasts XREF_BIBR - XREF_BIBR, though MITF is not expressed in the neural crest.

      WNT3A activates Wnt.

    41. Inhibition of Wnt signaling by a Wnt antagonist secreted frizzled related protein 4 (sFRP4), which is exclusively expressed in the epithelial cells but not the melanocytes of the hair follicle, results in a decrease of melanocytes differentiation in the regenerating hair follicle 79.

      SFRP4 activates Wnt.

    42. PDGF promotes the proliferation of human melanoblasts and differentiation of melanocytes 104, indicating that adipose secreted PDGF may also regulate McSCs activation and differentiation.
    43. Mechanistically, knockout of Kindlin-1 promotes cutaneous epithelial stem cells differentiation via inhibiting alpha (v) beta (6) integrin mediated TGF-beta1 liberation and promoting integrin independent Wnt ligand expression to activate Wnt and beta-catenin signaling 82.

      Integrins activates TGFB1.

    44. Injecting Wnt inhibitor DKK1 into skin inhibits TPA induced the proliferation and differentiation of McSCs 52.
    45. Injecting Wnt inhibitor DKK1 into skin inhibits TPA induced the proliferation and differentiation of McSCs 52.
    46. So the white color can be due to the absence of melanocytes by non migration or death of melanocytes, or due to the suppression of differentiation by agouti or other inhibitors.

      Agouti activates cell differentiation.

    47. HGF, SCF, and End3 have been revealed to promote melanoblast or melanocyte proliferation and differentiation XREF_BIBR, XREF_BIBR.

      End3 activates cell differentiation.

    48. HGF, SCF, and End3 have been revealed to promote melanoblast or melanocyte proliferation and differentiation XREF_BIBR, XREF_BIBR.
    1. In contrast, blockade of CD16 produced no inhibitory effect on CRP-induced expression of CXCL8, CCL2, MMP9 and IL-6 by RA-FLSs ( xref ), suggesting that CRP may not signal through the CD16 to induce joint inflammation in vitro .

      CRP increases the amount of MMP9.

    2. However, blocking CD32 but not CD64 to inhibit CRP induced FLS proliferation, invasiveness, and proinflammatory cytokine CXCL8 production revealed a major role for CD32 signaling in synovial inflammation, although CRP via CD64, not CD32, to induce MMP9 expression was noticed.

      CRP increases the amount of MMP9.

    3. Interestingly, CRP-induced MMP9 expression and invasion on RA-FLSs were p38-dependent as addition of a p38 inhibitor (SB202190) but not a NF-κB inhibitor (PDTC) was capable of inhibiting CRP-induced MMP9 expression and cell invasion.

      CRP increases the amount of MMP9.

    4. In contrast, blockade of CD16 produced no inhibitory effect on CRP-induced expression of CXCL8, CCL2, MMP9 and IL-6 by RA-FLSs ( xref ), suggesting that CRP may not signal through the CD16 to induce joint inflammation in vitro .

      CRP increases the amount of CXCL8.

    5. However, CRP- induced expression of CXCL8 was CD32-dependent as it was blunted by the antibody against CD32, whereas CRP-induced MMP9 was blocked by the antibody to CD64, demonstrating that differential signaling mechanisms for CRP in regulating CXCL8 and MMP9 expression in RA-FLSs.

      CRP increases the amount of CXCL8.

    6. In contrast, blockade of CD16 produced no inhibitory effect on CRP-induced expression of CXCL8, CCL2, MMP9 and IL-6 by RA-FLSs ( xref ), suggesting that CRP may not signal through the CD16 to induce joint inflammation in vitro .

      CRP increases the amount of IL6.

    7. As shown in xref , CRP-induced expression of CCL2 and IL-6 was blocked by either neutralizing antibody to CD32 or CD64 or both, suggesting that CRP signals through both CD32/CD64 to induce expression of CCL2 and IL-6.

      CRP increases the amount of IL6.

    8. As shown in XREF_FIG, CRP induced expression of CCL2 and IL-6 was blocked by either neutralizing antibody to CD32 or CD64 or both, suggesting that CRP signals through both CD32 and CD64 to induce expression of CCL2 and IL-6.

      CRP increases the amount of IL6.

    9. In contrast, blockade of CD16 produced no inhibitory effect on CRP-induced expression of CXCL8, CCL2, MMP9 and IL-6 by RA-FLSs ( xref ), suggesting that CRP may not signal through the CD16 to induce joint inflammation in vitro .

      CRP increases the amount of CCL2.

    10. As shown in xref , CRP-induced expression of CCL2 and IL-6 was blocked by either neutralizing antibody to CD32 or CD64 or both, suggesting that CRP signals through both CD32/CD64 to induce expression of CCL2 and IL-6.

      CRP increases the amount of CCL2.

    11. As shown in XREF_FIG, CRP induced expression of CCL2 and IL-6 was blocked by either neutralizing antibody to CD32 or CD64 or both, suggesting that CRP signals through both CD32 and CD64 to induce expression of CCL2 and IL-6.

      CRP increases the amount of CCL2.

    12. We thus blocked CD32 or CD64 or both with neutralizing antibodies to differentially determine the signaling mechanisms of CRP-induced cytokine expression.

      CRP increases the amount of cytokine.

    13. It has been reported that CRP-induced cytokine expression is also regulated by TWIST transcriptionally in myeloma cells ( xref ).

      CRP increases the amount of cytokine.

    14. Thus, it is highly possible that high concentration of CRP in synovial fluid in patients with RA may directly bind primarily CD32 to activate p38 MAP kinase and NF-kappaB signaling in RA-FLSs to differentially regulate synovial inflammation.

      CD32 binds CRP.

    15. In vitro studies confirmed this notion and found that CRP was able to upregulate both CD32 and CD64 and induced FLS proliferation, invasion, and pro inflammatory expression by increasing production of CCL2, CXCL8, IL-6, MMP2, MMP9 while suppressing an anti-inflammatory cytokine IL-10 expression.
    16. RA-FLS was a major cell type responsible for CRP production in RA patients, accounting for more than 65% of CRP producing cells as identified by co-expressing CRP and vimentin in the inflamed synovial tissues in patients with RA.
    17. As shown in XREF_FIG, multiplex cytokine assay kits assays showed that addition of CRP dose-dependently upregulated CCL2, CXCL8, IL-6, MMP2, MMP9 in RA-FLS but not in HFLS, although expression of IL-1beta and TNFalpha was not significantly changed (XREF_FIG).

      CRP activates MMP9.

    18. In vitro studies confirmed this notion and found that CRP was able to upregulate both CD32 and CD64 and induced FLS proliferation, invasion, and pro inflammatory expression by increasing production of CCL2, CXCL8, IL-6, MMP2, MMP9 while suppressing an anti-inflammatory cytokine IL-10 expression.

      CRP activates MMP9.

    19. In vitro studies confirmed this notion and found that CRP was able to upregulate both CD32 and CD64 and induced FLS proliferation, invasion, and pro inflammatory expression by increasing production of CCL2, CXCL8, IL-6, MMP2, MMP9 while suppressing an anti-inflammatory cytokine IL-10 expression.

      CRP activates MMP2.

    20. As shown in XREF_FIG, multiplex cytokine assay kits assays showed that addition of CRP dose-dependently upregulated CCL2, CXCL8, IL-6, MMP2, MMP9 in RA-FLS but not in HFLS, although expression of IL-1beta and TNFalpha was not significantly changed (XREF_FIG).

      CRP activates MMP2.

    21. In vitro studies confirmed this notion and found that CRP was able to upregulate both CD32 and CD64 and induced FLS proliferation, invasion, and pro inflammatory expression by increasing production of CCL2, CXCL8, IL-6, MMP2, MMP9 while suppressing an anti-inflammatory cytokine IL-10 expression.

      CRP activates CXCL8.

    22. As shown in XREF_FIG, multiplex cytokine assay kits assays showed that addition of CRP dose-dependently upregulated CCL2, CXCL8, IL-6, MMP2, MMP9 in RA-FLS but not in HFLS, although expression of IL-1beta and TNFalpha was not significantly changed (XREF_FIG).

      CRP activates CXCL8.

    23. In vitro studies confirmed this notion and found that CRP was able to upregulate both CD32 and CD64 and induced FLS proliferation, invasion, and pro inflammatory expression by increasing production of CCL2, CXCL8, IL-6, MMP2, MMP9 while suppressing an anti-inflammatory cytokine IL-10 expression.

      CRP activates IL6.

    24. As shown in XREF_FIG, multiplex cytokine assay kits assays showed that addition of CRP dose-dependently upregulated CCL2, CXCL8, IL-6, MMP2, MMP9 in RA-FLS but not in HFLS, although expression of IL-1beta and TNFalpha was not significantly changed (XREF_FIG).

      CRP activates IL6.

    25. In vitro studies confirmed this notion and found that CRP was able to upregulate both CD32 and CD64 and induced FLS proliferation, invasion, and pro inflammatory expression by increasing production of CCL2, CXCL8, IL-6, MMP2, MMP9 while suppressing an anti-inflammatory cytokine IL-10 expression.

      CRP activates CCL2.

    26. As shown in XREF_FIG, multiplex cytokine assay kits assays showed that addition of CRP dose-dependently upregulated CCL2, CXCL8, IL-6, MMP2, MMP9 in RA-FLS but not in HFLS, although expression of IL-1beta and TNFalpha was not significantly changed (XREF_FIG).

      CRP activates CCL2.

    27. In vitro studies confirmed this notion and found that CRP was able to upregulate both CD32 and CD64 and induced FLS proliferation, invasion, and pro inflammatory expression by increasing production of CCL2, CXCL8, IL-6, MMP2, MMP9 while suppressing an anti-inflammatory cytokine IL-10 expression.
    28. This was further confirmed by the ability of pre-treating RA-FLS with a NF-kappaB inhibitor, PDTC (100 mumol/L) to inhibit CRP induced proliferation (XREF_FIG) and upregulation of CXCL8, CCL2.
    29. Here we tested the hypothesis that CRP may be produced locally by FLSs and functions to induce the synovial inflammation in patients with RA.
    30. CRP may promote synovial inflammation via mechanism associated with activation of CD32/64- p38 and NF-kappaB signaling.
    31. In the present study, we found that CRP signaled primarily through CD32, to a less extent of CD64, to differentially regulate joint inflammation.
    32. CRP Promotes RA-FLS Pro inflammatory Response Differentially via the CD32/64-p 38 and NF-kappaB-Dependent Mechanisms in vitro.
    33. CRP can induce synovial inflammation via mechanisms associated with activation of CD32/64-p 38 and NF-kappaB signaling.
    34. This was supported by the findings that CRP induced activation of p38 MAP kinase and NF-kappaB signaling was blunted by neutralizing antibodies against CD32 but not CD64.

      CRP activates p38.

    35. To examine whether CRP induces NF-kappaB nuclear translation, immunofluorescence and subcellular fractionation were performed.

      CRP activates NFkappaB.

    1. We found that p-Y42 RhoA also readily translocated to the nucleus, similarly to β-catenin, upon Wnt3A signaling ( xref A).

      RHOA translocates to the nucleus.

    2. However, the mechanism by which p -Tyr42 RhoA translocates to the nucleus remains elusive.

      RHOA translocates to the nucleus.

    3. We found that p-Y42 RhoA also readily translocated to the nucleus, similarly to beta-catenin, upon Wnt3A signaling (XREF_FIG A).

      RHOA translocates to the nucleus.

    4. However, the mechanism by which p-Tyr42 RhoA translocates to the nucleus remains elusive.

      RHOA translocates to the nucleus.

    5. In absence of Wnt signaling, glycogen synthase kinase-3 beta (GSK-3beta) phosphorylates beta-catenin in a ' destruction complex '.

      GSK3B phosphorylates CTNNB1.

    6. In presence of Wnt, there is the stimulation of cells in the canonical pathway as GSK-3beta phosphorylates lipoprotein receptor related protein 6 (LRP6) rather than beta-catenin, which is necessary for Axin to bind to LRP6 together with GSK-3beta and APC [XREF_BIBR].

      GSK3B phosphorylates CTNNB1.

    7. Consequently, GSK-3beta can not interact with and phosphorylate beta-catenin, resulting in increased beta-catenin stability.

      GSK3B phosphorylates CTNNB1.

    8. In particular, Tyr42 of RhoA phosphorylated by c-Src binds to IKKgamma, leading to IKKbeta activation, IkappaB phosphorylation and its degradation, resulting in NF-kappaB activation and tumorigenesis [XREF_BIBR].

      SRC phosphorylates RHOA on Y42.

    9. These results suggest that upon Wnt3A stimulation, p-Tyr216 GSK-3beta and dephospho-Ser9 GSK-3beta, both of which are known to be active forms of GSK-3beta, are critical for beta-catenin accumulation along with Tyr416 phosphorylation of Src and Tyr42 phosphorylation of RhoA (XREF_FIG C).

      SRC phosphorylates RHOA on Y42.

    10. In a previous report, we found that Tyr42 phosphorylation of RhoA (p-Tyr42 RhoA) is induced by hydrogen peroxide (H 2 O 2) in RAW264.7 cells, a macrophage cell line [XREF_BIBR].

      hydrogen peroxide leads to the phosphorylation of RHOA on Y42.

    11. Here, hydrogen peroxide also induced Tyr42 phosphorylation of Rho GTPase in HEK293T cells (XREF_FIG A).

      hydrogen peroxide leads to the phosphorylation of RHOA on Y42.

    12. Likewise, si-RhoA also prevented beta-catenin activity, and reconstitution of RhoA WT and phospho-mimetic Y42E restored the beta-catenin activity, but not with dephospho-mimetic RhoA Y42F, it having a remarkably reduced beta-catenin activity (XREF_FIG F).

      RHOA inhibits CTNNB1.

    13. However, in consistent to the previous report [XREF_BIBR], we revealed that si-RhoA impaired beta-catenin accumulation and reconstitution of RhoA restored beta-catenin accumulation (XREF_FIG A), suggesting that RhoA activity and beta-catenin stability are very closely linked, although the underlying mechanism by which RhoA induces beta-catenin accumulation in response to Wnt3A remains to be revealed.

      RHOA inhibits CTNNB1.

    14. Conversely, RhoA can be inactivated by GTPase activating proteins (GAPs) which catalyze GTP hydrolysis, leading to a GDP bound RhoA, which is inactive.

      GTP inhibits RHOA.

    15. RhoA WT and RhoA Y42E transfection to cells also had increased expression of cyclin D1 and c-Myc in presence of Wnt3A, whereas RhoA Y42F transfection inhibited this cyclin D1 and c-Myc up-regulation (XREF_FIG F).

      RHOA increases the amount of MYC.

    16. RhoA WT and RhoA Y42E transfection to cells also had increased expression of cyclin D1 and c-Myc in presence of Wnt3A, whereas RhoA Y42F transfection inhibited this cyclin D1 and c-Myc up-regulation (XREF_FIG F).

      RHOA increases the amount of Cyclin.

    17. In this study, we found that Wnt3A induces interaction of p-Tyr42 RhoA and beta-catenin and p-Tyr42 RhoA delivers beta-catenin to the nucleus, where p-Tyr42 RhoA as well as beta-catenin regulates expression of specific genes such as Vim by binding to Vim promoter.

      CTNNB1 binds RHOA.

    18. We next tried to identify whether GDP/GTP bound states of RhoA determine the interaction between RhoA and beta-catenin.

      CTNNB1 binds RHOA.

    19. In addition, it was demonstrated that p-Tyr42 RhoA directly interact with beta-catenin and nuclear localization of p-Tyr42 RhoA is required for nuclear delivery of beta-catenin.

      CTNNB1 binds RHOA.

    20. We next tried to identify whether GDP/GTP-bound states of RhoA determine the interaction between RhoA and β-catenin.

      CTNNB1 binds RHOA.

    21. Recombinant GST-β-catenin binding to RhoA-GTPγS or RhoA-GDP in vitro was not significantly different, suggesting that GDP or GTP binding to RhoA is not critical for regulating interaction between RhoA with β-catenin ( xref D).

      CTNNB1 binds RHOA.

    22. To identify the specific domain of β-catenin associating with RhoA, we expressed and purified GST-fusions of β-catenin domains composed of amino acids (aa) 1–140, 141–390, 391–662 and 663–782 ( xref E and F).

      CTNNB1 binds RHOA.

    23. Similarly, GST-RhoA phosphorylated by Src and ATP [ xref ] also readily bound to β-catenin in vitro , irrespective of GDP- or GTPγS-preloaded RhoA, suggesting that phosphorylation of Tyr42, but not GTP/GDP-binding state is critical for interaction between RhoA and β-catenin ( xref D).

      CTNNB1 binds RHOA.

    24. The 3D of RhoA revealed Tyr42 residue positioning in extended β2 region did not dramatically alter 3D location, while Tyr34 in switch 1 (aa 28–38) and Tyr66 in switch 2 (aa 61–78) regions revealed significant alteration [ xref ] ( xref E), suggesting that GDP or GTP may not contribute p -Tyr42 RhoA binding to β-catenin.

      CTNNB1 binds RHOA.

    25. Interaction of p -Tyr42 RhoA with β-catenin.

      CTNNB1 binds RHOA.

    26. Also, RhoA preferentially binds to N -terminal domain (NTD: aa 1–140) of β-catenin ( xref G).

      CTNNB1 binds RHOA.

    27. It is remarkable that RhoA Y42F (dephospho-mimetic) did not interfere with β-catenin accumulation ( xref A), but RhoA Y42F could not bind to β-catenin ( xref B and 5C).

      CTNNB1 binds RHOA.

    28. Since expression of vimentin can be also induced by β-catenin [ xref ], we speculate p -Tyr42 RhoA binding to β-catenin facilitate vimentin expression.

      CTNNB1 binds RHOA.

    29. Meanwhile, p -Tyr42 RhoA bound to β-catenin via the N -terminal domain of β-catenin, thereby leading to the nuclear translocation of p -Tyr42 RhoA/β-catenin complex.

      CTNNB1 binds RHOA.

    30. In this study, we found that Wnt3A induces interaction of p -Tyr42 RhoA and β-catenin and p -Tyr42 RhoA delivers β-catenin to the nucleus, where p -Tyr42 RhoA as well as β-catenin regulates expression of specific genes such as Vim by binding to Vim promoter.

      CTNNB1 binds RHOA.

    31. In addition, it was demonstrated that p -Tyr42 RhoA directly interact with β-catenin and nuclear localization of p -Tyr42 RhoA is required for nuclear delivery of β-catenin.

      CTNNB1 binds RHOA.

    32. Of note, beta-catenin binds to Lef1 and activates Lef1 transcription complex, leading to down-regulation of E-cadherin expression [XREF_BIBR].

      CTNNB1 binds LEF1.

    33. Of note, β-catenin binds to Lef1 and activates Lef1 transcription complex, leading to down-regulation of E -cadherin expression [ xref ].

      CTNNB1 binds LEF1.

    34. p -Tyr42 Rho binds to promoters of specific genes such as Vim.

      VIM binds RHOA.

    35. In this study, as a novel mechanism, p -Tyr42 RhoA binds to promoter of Vim and regulates vimentin expression, suggesting that p -Tyr42 RhoA regulates EMT ( xref ).

      VIM binds RHOA.

    36. Notably, p -Tyr42 RhoA as well as β-catenin was associated with the promoter of Vim , leading to increased expression of vimentin.

      VIM binds CTNNB1 and RHOA.

    37. ROCK2 and p-Tyr42 RhoA bind to Src, where they transmit other signaling pathway through p-p47phox (XREF_FIG H, XREF_FIG and 3L).

      SRC binds RHOA.

    38. Notably, p-Tyr42 RhoA preferentially bound to ROCK2, which in turn preferentially bound to p47phox, leading Ser345 phosphorylation of p47phox (XREF_FIG H and XREF_FIG).

      ROCK2 binds RHOA.

    39. Notably, p -Tyr42 RhoA preferentially bound to ROCK2, which in turn preferentially bound to p47phox, leading Ser345 phosphorylation of p47phox ( xref H and xref ).

      ROCK2 binds RHOA.

    40. It is notable that IKKgamma (also known as NEMO) facilitates RhoA activation via IKKgamma (NEMO) causing the dissociation of the RhoA and RhoGDI complex [XREF_BIBR].

      RhoGDI binds RHOA.

    41. It is notable that IKKγ (also known as NEMO) facilitates RhoA activation via IKKγ (NEMO) causing the dissociation of the RhoA-RhoGDI complex [ xref ].

      RhoGDI binds RHOA.

    42. GST-RhoA WT, Y42E and Y42F preloaded with GDP or GTPγS were incubated recombinant purified β-catenin in vitro .

      GST binds RHOA.

    43. Similarly, GST-RhoA phosphorylated by Src and ATP [ xref ] also readily bound to β-catenin in vitro , irrespective of GDP- or GTPγS-preloaded RhoA, suggesting that phosphorylation of Tyr42, but not GTP/GDP-binding state is critical for interaction between RhoA and β-catenin ( xref D).

      GST binds RHOA.

    44. Recombinant GST-β-catenin binding to RhoA-GTPγS or RhoA-GDP in vitro was not significantly different, suggesting that GDP or GTP binding to RhoA is not critical for regulating interaction between RhoA with β-catenin ( xref D).

      GST binds CTNNB1.

    45. RhoA preloaded with GTPγS readily bound to aa 1–140 NTD of GST-β-catenin domain, conjugated to beads, whereas other domain fusion proteins only marginally bound to RhoA ( xref G).

      GST binds CTNNB1.

    46. Conversely, RhoA can be inactivated by GTPase activating proteins (GAPs) which catalyze GTP hydrolysis, leading to a GDP bound RhoA, which is inactive.

      GDP binds RHOA.

    47. As a small GTPase, RhoA can be activated by several guanine nucleotide exchange factors (GEFs) resulting in GTP binding to RhoA, representing an activated RhoA.

      GTP binds RHOA.

    48. Remarkably, si-RhoA impaired beta-catenin accumulation, but re-constitutive dephospho-mimetic RhoA Y42F as well as RhoA WT and phospho-mimetic RhoA Y42E allowed recovery of beta-catenin accumulation (XREF_FIG A).

      RHOA activates CTNNB1.

    49. However, in consistent to the previous report [XREF_BIBR], we revealed that si-RhoA impaired beta-catenin accumulation and reconstitution of RhoA restored beta-catenin accumulation (XREF_FIG A), suggesting that RhoA activity and beta-catenin stability are very closely linked, although the underlying mechanism by which RhoA induces beta-catenin accumulation in response to Wnt3A remains to be revealed.

      RHOA activates CTNNB1.

    50. One clue for this function is that the RhoA effector protein, ROCK2, which was supposed to be activated by p-Tyr42 RhoA (XREF_FIG H), may enhance the activation of p300 acetyltransferase by phosphorylation [XREF_BIBR].

      RHOA activates ROCK2.

    51. The results suggest that ROCK2 is a preferential downstream effector protein of p-Tyr42 RhoA while ROCK1 is activated dominantly by RhoA (XREF_FIG J).

      RHOA activates ROCK1.

    52. The results suggest that ROCK2 is a preferential downstream effector protein of p -Tyr42 RhoA while ROCK1 is activated dominantly by RhoA ( xref J).

      RHOA activates ROCK1.

    53. Consistent to the result of XREF_FIG B, si-RhoA abolished superoxide production, and reconstituted RhoA WT and phospho-mimetic RhoA Y42E restored superoxide production.

      RHOA activates superoxide.

    54. Of note, β-catenin binds to Lef1 and activates Lef1 transcription complex, leading to down-regulation of E -cadherin expression [ xref ].

      CTNNB1 activates LEF1.

    55. 3.4 Wnt3A induces formation of RhoA, beta, and catenin complex.

      WNT3A activates RHOA.

    56. Here, we found that Wnt3A also induces the complex formation of beta-catenin and RhoA, particularly p-Y42 RhoA.

      WNT3A activates RHOA.

    57. Wnt3A induced nuclear localization of beta-catenin (XREF_FIG A).

      WNT3A activates CTNNB1.

    58. Here, we found that Wnt3A also induces the complex formation of beta-catenin and RhoA, particularly p-Y42 RhoA.

      WNT3A activates CTNNB1.

    59. Of note, vimentin can activate RhoA through GEF-H1 (also known as ARHGEF2) [ xref ].

      VIM activates RHOA.

    60. Of note, vimentin can activate RhoA through GEF-H1 (also known as ARHGEF2) [XREF_BIBR].

      VIM activates RHOA.

    61. ROS scavenger, NAC and NADPH oxidase inhibitors, DPI and apocynin abolished upregulation of p-Tyr42 Rho, beta-catenin, p-Tyr416 Src, p-Ser9 GSK-3beta and p-Tyr216 GSK-3beta upon Wnt3A in HEK293T cells, suggesting that superoxide is closely involved in the increases of the above molecules via Wnt3A (XREF_FIG C).
    62. ROS scavenger, NAC and NADPH oxidase inhibitors, DPI and apocynin abolished upregulation of p-Tyr42 Rho, beta-catenin, p-Tyr416 Src, p-Ser9 GSK-3beta and p-Tyr216 GSK-3beta upon Wnt3A in HEK293T cells, suggesting that superoxide is closely involved in the increases of the above molecules via Wnt3A (XREF_FIG C).
    63. As a small GTPase, RhoA can be activated by several guanine nucleotide exchange factors (GEFs) resulting in GTP binding to RhoA, representing an activated RhoA.

      guanine activates RHOA.

    64. Thereby, it is possible that p -Tyr42 RhoA GTPase causes the acetylation of β-catenin through p300 acetyltransferase, remarkably enhancing β-catenin activity as a co-activator of TCF4 in the nucleus.

      RHOA leads to the acetylation of CTNNB1.

    65. Thereby, it is possible that p-Tyr42 RhoA GTPase causes the acetylation of beta-catenin through p300 acetyltransferase, remarkably enhancing beta-catenin activity as a co-activator of TCF4 in the nucleus.

      RHOA leads to the acetylation of CTNNB1.

    1. RhoA GTPase phosphorylated at tyrosine 42 by src kinase binds to β-catenin and contributes transcriptional regulation of vimentin upon Wnt3A.

      CTNNB1 binds RHOA.

    2. RhoA GTPase phosphorylated at tyrosine 42 by src kinase binds to beta-catenin and contributes transcriptional regulation of vimentin upon Wnt3A.

      CTNNB1 binds RHOA.

    3. Meanwhile, p-Tyr42 RhoA bound to β-catenin via the N-terminal domain of β-catenin, thereby leading to the nuclear translocation of p-Tyr42 RhoA/β-catenin complex.

      CTNNB1 binds RHOA.

    4. Meanwhile, p-Tyr42 RhoA bound to beta-catenin via the N-terminal domain of beta-catenin, thereby leading to the nuclear translocation of p-Tyr42 RhoA, beta, and catenin complex.

      CTNNB1 binds RHOA.

    5. Notably, p-Tyr42 RhoA as well as β-catenin was associated with the promoter of Vim, leading to increased expression of vimentin.

      VIM binds CTNNB1 and RHOA.

    6. RhoA GTPase phosphorylated at tyrosine 42 by src kinase binds to beta-catenin and contributes transcriptional regulation of vimentin upon Wnt3A.

      RHOA activates VIM.

    7. Wnt3A also induced accumulation of beta-catenin along with activations of RhoA and ROCK1.

      WNT3A activates CTNNB1.

    1. KLK8 degrades VE-cadherin , thereby promoting plakoglobin nuclear translocation The KLK family is considered to cleave kininogens to release kinin 15 .

      KLK8 inhibits CDH5.

    2. KLK8 degrades VE-cadherin, thereby promoting plakoglobin nuclear translocation.

      KLK8 inhibits CDH5.

    3. As shown in supplemental xref , treatment of cardiac fibroblasts with Ad-KLK8 induced mRNA expression of the proliferation-related genes Ki67, proliferating cell nuclear antigen (PCNA) and cyclin D1 xref .

      KLK8 increases the amount of MKI67.

    4. Protein expression levels of Ki67 and PCNA were also increased by KLK8 overexpression.

      KLK8 increases the amount of MKI67.

    5. The HIF-1α inhibitor echinomycin not only inhibited basal TGF-β1 expression, but also blocked KLK8-induced TGF-β1 mRNA expression and release in HCAECs (Figure xref D).

      KLK8 increases the amount of TGFB1.

    6. However, ICG-001 treatment had no significant effect on KLK8-induced TGF-β1 expression ( xref F).

      KLK8 increases the amount of TGFB1.

    7. The present study found that upregulation of KLK8 leads to plakoglobin dependent nuclear translocation of p53, which binds to HIF-1alpha, and further enhances the transactivation effect of HIF-1alpha on the TGF-beta1 promoter and consequently upregulates TGF-beta1 expression at the transcriptional level.

      KLK8 increases the amount of TGFB1.

    8. As shown in supplemental xref , treatment of cardiac fibroblasts with Ad-KLK8 induced mRNA expression of the proliferation-related genes Ki67, proliferating cell nuclear antigen (PCNA) and cyclin D1 xref .

      KLK8 increases the amount of messenger RNA.

    9. The HIF-1α inhibitor echinomycin not only inhibited basal TGF-β1 expression, but also blocked KLK8-induced TGF-β1 mRNA expression and release in HCAECs (Figure xref D).

      KLK8 increases the amount of messenger RNA.

    10. A specificity protein-1 (Sp-1) consensus site was identified in the human KLK8 promoter and was found to mediate the high glucose induced KLK8 expression.

      SP1 increases the amount of KLK8.

    11. A specificity protein-1 (Sp-1) consensus site was identified in the human KLK8 promoter and was found to mediate the high glucose induced KLK8 expression.

      SP1 increases the amount of KLK8.

    12. It was found that the mRNA and protein expression levels of KLK8 were significantly upregulated in a dose dependent manner in high glucose treated HCAECs compared with those of normal glucose treated HCAECs, suggesting that high glucose stimulated KLK8 expression at the transcriptional level.

      glucose increases the amount of KLK8.

    13. To explore the molecular mechanisms involved in the high glucose induced upregulation of KLK8 expression, HCAECs were treated with increasing concentration of glucose (15 and 25 mM) for 5 days.

      glucose increases the amount of KLK8.

    14. The present study found that Ad-KLK8 infection resulted in a 2.88 ± 0.35 fold increase in the content of bradykinin in the culture medium of HCAECs.

      KLK8 binds Ad.

    15. In HCAECs, it was found that Ad-KLK8 increased the expression levels of α-SMA and vimentin, whereas it decreased the expression levels of CD31 and VE-cadherin, in a dose-dependent manner, suggesting that KLK8 overexpression is able to induce EndMT (Figure xref E-F).

      KLK8 binds Ad.

    16. In addition, the permeability of a confluent HCAEC monolayer was measured for 10-kDa FITC-dextran, and it was found that Ad-KLK8 treatment significantly increased endothelial permeability (Figure xref L).

      KLK8 binds Ad.

    17. Infection of HCAECs with increasing concentrations of KLK8 adenovirus (Ad-KLK8) led to an increase in KLK8 expression in a dose-dependent manner ( xref ).

      KLK8 binds Ad.

    18. As shown in supplemental xref , treatment of cardiac fibroblasts with Ad-KLK8 induced mRNA expression of the proliferation-related genes Ki67, proliferating cell nuclear antigen (PCNA) and cyclin D1 xref .

      KLK8 binds Ad.

    19. The present study then examined the composition of the endothelial cell culture medium after cell transfection with Ad-KLK8.

      KLK8 binds Ad.

    20. Adenovirus-mediated overexpression of KLK8 (Ad-KLK8) resulted in increases in endothelial cell damage, permeability and transforming growth factor (TGF)-β1 release in HCAECs.

      KLK8 binds Ad.

    21. The present study found that KLK8 overexpression significantly increased the association of p53 with Smad3, which was blocked by plakoglobin knockdown (Figure xref F).

      TP53 binds SMAD3.

    22. KLK8 also induced the binding of p53 with Smad3, subsequently promoting pro-EndMT reprogramming via the TGF-β1/Smad signaling pathway in HCAECs.

      TP53 binds SMAD3.

    23. KLK8 also induced the binding of p53 with Smad3, subsequently promoting pro EndMT reprogramming via the TGF-beta1 and Smad signaling pathway in HCAECs.

      TP53 binds SMAD3.

    24. Previous study has found that plakoglobin interacts with p53 and increases its transcriptional activity, thereby regulating the expression of various genes involved in tumorigenesis and metastasis xref .

      TP53 binds JUP.

    25. The association of plakoglobin with p53 in diabetic heart tissues was decreased in KLK8 -/- mice (Figure xref F).

      TP53 binds JUP.