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active rac1 pak 02  (Cytoskeleton Inc)


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    Cytoskeleton Inc active rac1 pak 02
    a , Immunoblots showing cleaved Notch1 (c-Notch1) and total Notch1 in MVECs treated with S1P for 1-hour ± γ-secretase inhibitor DAPT; quantification below (mean ± SEM). b , Fluorescent micrographs of YFP-Notch1 reporter intensity after S1P treatment; quantification below. Scale bar, 50 μm. c-d , Fold change in HES1 and HEY1 gene expression levels in (c) MVECs after S1P ± DAPT treatment, and in (d) mouse tissues after intravenous injection of S1P ± DAPT. e , Evans blue dye leakage in mouse dermis 1-hour post-injection with DMSO, S1P, or S1P + DAPT. Scale bar, 5 mm. f-i , Quantified dye leakage in (f) dermis, (g) lung, (h) liver, and (i) kidney, measured via absorbance. j , Heat maps of dextran dye diffusion in engineered microvessels ± S1P. Scale bar, 50 μm. k-l , Diffusive permeability in microvessels lined with (k) DAPT-vs. DMSO-treated or (l) NOTCH1 KO vs. SCR KO MVECs (mean ± SD). m , Micrographs of VE-cadherin (magenta), actin (green), and nuclei (blue) staining in microvessels. Scale bar, 50 μm. n-o , Mean corrected intensities of junctional (n) VE-cadherin and (o) actin from (m). p , <t>Rac1</t> activity (PBD pull-down). Active/total Rac1 quantification below. q , Schematic of canonical Notch signaling. r , HES1 / HEY1 gene expression levels in dominant negative-MAML-GFP (DN-MAML-GFP) normalized to GFP expressing cells. s , Permeability in DN-MAML-GFP vs GFP-lined microvessels ± S1P. t , Schematic of Notch cortical pathway mediated by its transmembrane domain (TMD). u , Permeability in TMD-RFP vs RFP-lined microvessels ± S1P. v , Permeability in TMD-RFP vs RFP-lined microvessels ± clinical S1P receptor degrader fingolimod.
    Active Rac1 Pak 02, supplied by Cytoskeleton Inc, used in various techniques. Bioz Stars score: 95/100, based on 161 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pak+pbd+beads/PAK-PBD+beads+-+binds+active+Rac+Cdc42+proteins/bio_rxiv__64898__2026__05__20__726610-282-4-10
    Average 95 stars, based on 161 article reviews
    active rac1 pak 02 - by Bioz Stars, 2026-10
    95/100 stars

    Images

    1) Product Images from "Sphingosine-1-phosphate cross-talks to Notch via a S1PR1-Dll4-MPDZ complex to regulate endothelial barrier function"

    Article Title: Sphingosine-1-phosphate cross-talks to Notch via a S1PR1-Dll4-MPDZ complex to regulate endothelial barrier function

    Journal: bioRxiv

    doi: 10.64898/2026.05.20.726610

    a , Immunoblots showing cleaved Notch1 (c-Notch1) and total Notch1 in MVECs treated with S1P for 1-hour ± γ-secretase inhibitor DAPT; quantification below (mean ± SEM). b , Fluorescent micrographs of YFP-Notch1 reporter intensity after S1P treatment; quantification below. Scale bar, 50 μm. c-d , Fold change in HES1 and HEY1 gene expression levels in (c) MVECs after S1P ± DAPT treatment, and in (d) mouse tissues after intravenous injection of S1P ± DAPT. e , Evans blue dye leakage in mouse dermis 1-hour post-injection with DMSO, S1P, or S1P + DAPT. Scale bar, 5 mm. f-i , Quantified dye leakage in (f) dermis, (g) lung, (h) liver, and (i) kidney, measured via absorbance. j , Heat maps of dextran dye diffusion in engineered microvessels ± S1P. Scale bar, 50 μm. k-l , Diffusive permeability in microvessels lined with (k) DAPT-vs. DMSO-treated or (l) NOTCH1 KO vs. SCR KO MVECs (mean ± SD). m , Micrographs of VE-cadherin (magenta), actin (green), and nuclei (blue) staining in microvessels. Scale bar, 50 μm. n-o , Mean corrected intensities of junctional (n) VE-cadherin and (o) actin from (m). p , Rac1 activity (PBD pull-down). Active/total Rac1 quantification below. q , Schematic of canonical Notch signaling. r , HES1 / HEY1 gene expression levels in dominant negative-MAML-GFP (DN-MAML-GFP) normalized to GFP expressing cells. s , Permeability in DN-MAML-GFP vs GFP-lined microvessels ± S1P. t , Schematic of Notch cortical pathway mediated by its transmembrane domain (TMD). u , Permeability in TMD-RFP vs RFP-lined microvessels ± S1P. v , Permeability in TMD-RFP vs RFP-lined microvessels ± clinical S1P receptor degrader fingolimod.
    Figure Legend Snippet: a , Immunoblots showing cleaved Notch1 (c-Notch1) and total Notch1 in MVECs treated with S1P for 1-hour ± γ-secretase inhibitor DAPT; quantification below (mean ± SEM). b , Fluorescent micrographs of YFP-Notch1 reporter intensity after S1P treatment; quantification below. Scale bar, 50 μm. c-d , Fold change in HES1 and HEY1 gene expression levels in (c) MVECs after S1P ± DAPT treatment, and in (d) mouse tissues after intravenous injection of S1P ± DAPT. e , Evans blue dye leakage in mouse dermis 1-hour post-injection with DMSO, S1P, or S1P + DAPT. Scale bar, 5 mm. f-i , Quantified dye leakage in (f) dermis, (g) lung, (h) liver, and (i) kidney, measured via absorbance. j , Heat maps of dextran dye diffusion in engineered microvessels ± S1P. Scale bar, 50 μm. k-l , Diffusive permeability in microvessels lined with (k) DAPT-vs. DMSO-treated or (l) NOTCH1 KO vs. SCR KO MVECs (mean ± SD). m , Micrographs of VE-cadherin (magenta), actin (green), and nuclei (blue) staining in microvessels. Scale bar, 50 μm. n-o , Mean corrected intensities of junctional (n) VE-cadherin and (o) actin from (m). p , Rac1 activity (PBD pull-down). Active/total Rac1 quantification below. q , Schematic of canonical Notch signaling. r , HES1 / HEY1 gene expression levels in dominant negative-MAML-GFP (DN-MAML-GFP) normalized to GFP expressing cells. s , Permeability in DN-MAML-GFP vs GFP-lined microvessels ± S1P. t , Schematic of Notch cortical pathway mediated by its transmembrane domain (TMD). u , Permeability in TMD-RFP vs RFP-lined microvessels ± S1P. v , Permeability in TMD-RFP vs RFP-lined microvessels ± clinical S1P receptor degrader fingolimod.

    Techniques Used: Western Blot, Gene Expression, Injection, Diffusion-based Assay, Permeability, Staining, Activity Assay, Dominant Negative Mutation, Expressing

    Related Articles

    Incubation:

    Article Title: Gut commensal metabolite rhamnose promotes macrophages phagocytosis by activating SLC12A4 and protects against sepsis in mice
    Article Snippet: Total protein was extracted from the cells with cell lysis buffer (Part# CLB01, Cytoskeleton, USA) containing Protease Inhibitor Cocktail (Cat# PIC02, Cytoskeleton, USA) and extracted protein was quantified with a PierceTM BCA protein assay kit (Cat# 23225, Thermo Fisher Scientific, USA). .. Proteins (300–800 μg) from the cell lysates were collected into each tube and added with 10 μg of PAK-PBD beads (Cat# PAK02, Cytoskeleton, USA) and then incubated at 4 °C on a rotator for 1 h. After centrifugation at 5000×g at 4 °C for 1 min, beads were washed once with a wash buffer (Cat# WB01, Cytoskeleton, USA), GTP-Rac1 and GTP-Cdc42 were eluted with 30 L of 2× loading buffer (Cat# FD006, Fdbio, China) and boiling for 5 min. Extracted protein was used for Western blot analysis. .. For phosphorylated SLC12A4 (p-SLC12A4) assay, BMDMs were treated with rhamnose (100 μmol/L) for 5 min and then cells were lysed in RIPA buffer (Cat# P0013B, Beyotime, China).

    Article Title: KLF7 promotes neuroblastoma differentiation through regulation neuroblast differentiation-associated protein AHNAKs and is a marker of clinical outcome
    Article Snippet: .. Cleared lysates were incubated for 1hr at 4°C with PAK-PBD beads (Cytoskeleton Inc.) to precipitate GTP-bound Rac1 and Cdc42, respectively. ..


    Article Title: KLF7 promotes neuroblastoma differentiation through the GTPase signaling pathway by upregulating neuroblast differentiation-associated protein AHNAKs and glycerophosphodiesterase GDPD5.
    Article Snippet: doi:10.1111/febs.17208 The arrest of neural crest-derived sympathoadrenal neuroblast differentiation contributes to neuroblastoma formation, and overriding this blocked differentiation is a clear strategy for treating high-risk neuroblastoma.. A better understanding of neuroblast or neuroblastoma differentiation is essential for developing new therapeutic approaches.. It has been proposed that Krueppel-like factor 7 (KLF7) is a neuroblastoma super-enhancerassociated transcription factor gene.

    Article Title: EPS8L1 promotes migration and metastasis of ovarian cancer by activating Rac1/MAPK signaling pathway via upregulating TIAM2
    Article Snippet: .. The lysates were centrifuged (10,000× g, 4 °C for 1 min) to remove cellular debris and then incubated with 10 μL of PAK-PBD beads from Rac1 Activation Kit (cytoskeleton, CAT#BK035-S) for 60 min at 4°C on a shaker. ..

    Article Title: LUZP1 Regulates Dendritic Spine Maturation and Synaptic Plasticity in the Hippocampal Dentate Gyrus of Mice
    Article Snippet: Research Articles | Development/Plasticity/Repair LUZP1 Regulates Dendritic Spine Maturation and Synaptic Plasticity in the Hippocampal Dentate Gyrus of Mice https://doi.org/10.1523/JNEUROSCI.1867-24.2025 Received: 30 September 2024 Revised: 16 February 2025 Accepted: 18 March 2025 Copyright © 2025 the authors This Early Release article has been peer reviewed and accepted, but has not been through the composition and copyediting processes.The final version may differ slightly in style or formatting and will contain links to any extended data.. Alerts: Sign up at www.jneurosci.org/alerts to receive customized email alerts when the fully formatted version of this article is published.. - 1 - LUZP1 Regulates Dendritic Spine Maturation and 1 Synaptic Plasticity in the Hippocampal Dentate Gyrus of 2 Mice 3 Abbreviated title: LUZP1 in Hippocampal Synaptic Plasticity 4 Xiaojie Wang 1,2,* , Liang Wang 1,* , Qian Bu 1, Yuzhou Xiao1, Yue Zhao1, Linhong Jiang1, 5 Yanping Dai 1 , Hongchun Li 1 , Haxiaoyu Liu 1 , Yaxing Chen 1 , Angelo D. Flores 2 , Yinglan Zhao 1 , 6 Xiaobo Cen 1,# 7 1.

    Centrifugation:

    Article Title: Gut commensal metabolite rhamnose promotes macrophages phagocytosis by activating SLC12A4 and protects against sepsis in mice
    Article Snippet: Total protein was extracted from the cells with cell lysis buffer (Part# CLB01, Cytoskeleton, USA) containing Protease Inhibitor Cocktail (Cat# PIC02, Cytoskeleton, USA) and extracted protein was quantified with a PierceTM BCA protein assay kit (Cat# 23225, Thermo Fisher Scientific, USA). .. Proteins (300–800 μg) from the cell lysates were collected into each tube and added with 10 μg of PAK-PBD beads (Cat# PAK02, Cytoskeleton, USA) and then incubated at 4 °C on a rotator for 1 h. After centrifugation at 5000×g at 4 °C for 1 min, beads were washed once with a wash buffer (Cat# WB01, Cytoskeleton, USA), GTP-Rac1 and GTP-Cdc42 were eluted with 30 L of 2× loading buffer (Cat# FD006, Fdbio, China) and boiling for 5 min. Extracted protein was used for Western blot analysis. .. For phosphorylated SLC12A4 (p-SLC12A4) assay, BMDMs were treated with rhamnose (100 μmol/L) for 5 min and then cells were lysed in RIPA buffer (Cat# P0013B, Beyotime, China).

    Western Blot:

    Article Title: Gut commensal metabolite rhamnose promotes macrophages phagocytosis by activating SLC12A4 and protects against sepsis in mice
    Article Snippet: Total protein was extracted from the cells with cell lysis buffer (Part# CLB01, Cytoskeleton, USA) containing Protease Inhibitor Cocktail (Cat# PIC02, Cytoskeleton, USA) and extracted protein was quantified with a PierceTM BCA protein assay kit (Cat# 23225, Thermo Fisher Scientific, USA). .. Proteins (300–800 μg) from the cell lysates were collected into each tube and added with 10 μg of PAK-PBD beads (Cat# PAK02, Cytoskeleton, USA) and then incubated at 4 °C on a rotator for 1 h. After centrifugation at 5000×g at 4 °C for 1 min, beads were washed once with a wash buffer (Cat# WB01, Cytoskeleton, USA), GTP-Rac1 and GTP-Cdc42 were eluted with 30 L of 2× loading buffer (Cat# FD006, Fdbio, China) and boiling for 5 min. Extracted protein was used for Western blot analysis. .. For phosphorylated SLC12A4 (p-SLC12A4) assay, BMDMs were treated with rhamnose (100 μmol/L) for 5 min and then cells were lysed in RIPA buffer (Cat# P0013B, Beyotime, China).

    Activation Assay:

    Article Title: EPS8L1 promotes migration and metastasis of ovarian cancer by activating Rac1/MAPK signaling pathway via upregulating TIAM2
    Article Snippet: .. The lysates were centrifuged (10,000× g, 4 °C for 1 min) to remove cellular debris and then incubated with 10 μL of PAK-PBD beads from Rac1 Activation Kit (cytoskeleton, CAT#BK035-S) for 60 min at 4°C on a shaker. ..

    Article Title: The overexpression of EPS8L1 upregulates TIAM2 to promote cytoskeleton remodeling by activating the Rac1/MAPK signaling pathway in the migration of ovarian cancer
    Article Snippet: .. The lysates were centrifuged (10,000× g, 4 °C for 1 min) to remove cellular debris and then incubated with 10 μL of PAK-PBD beads from a Rac1 Activation Kit (cytoskeleton, CAT#BK035-S) for 60 min at 4°C on a shaker. ..

    other:

    Article Title: The TCR-SUB1-DOCK2 axis promotes autoimmunity by driving pathogenic CD4 + T cell tissue infiltration.
    Article Snippet: Article The TCR-SUB1-DOCK2 axis promotes autoimmunity by driving pathogenic CD4+ T cell tissue infiltration Highlights • SUB1 dictates the tissue infiltration of pathogenic CD4+ T cells • Sub1 expression is directly modulated by the TCR-IRF4 pathway • Sub1 deficiency prevents EAE onset by halving DOCK2 levels • SUB1 undergoes LLPS to open chromatin and drive the JUNB-DOCK2 transcriptional cascade Authors Xiaoxue Li, Wenhua Liang, Weifang Wang, Eilon Sherman, Keling Huang, Feng Wang Correspondence wangfeng16@sjtu.edu.cn In brief Blocking pathogenic T cell migration is crucial for treating autoimmune diseases.. Li et al. reveal SUB1 as a gatekeeper of CD4+ T cell infiltration in autoimmunity.. TCR-induced SUB1 drives Rac- dependent cytoskeletal remodeling via DOCK2 transcriptional activation.

    Activity Assay:

    Article Title: LUZP1 Regulates Dendritic Spine Maturation and Synaptic Plasticity in the Hippocampal Dentate Gyrus of Mice
    Article Snippet: Research Articles | Development/Plasticity/Repair LUZP1 Regulates Dendritic Spine Maturation and Synaptic Plasticity in the Hippocampal Dentate Gyrus of Mice https://doi.org/10.1523/JNEUROSCI.1867-24.2025 Received: 30 September 2024 Revised: 16 February 2025 Accepted: 18 March 2025 Copyright © 2025 the authors This Early Release article has been peer reviewed and accepted, but has not been through the composition and copyediting processes.The final version may differ slightly in style or formatting and will contain links to any extended data.. Alerts: Sign up at www.jneurosci.org/alerts to receive customized email alerts when the fully formatted version of this article is published.. - 1 - LUZP1 Regulates Dendritic Spine Maturation and 1 Synaptic Plasticity in the Hippocampal Dentate Gyrus of 2 Mice 3 Abbreviated title: LUZP1 in Hippocampal Synaptic Plasticity 4 Xiaojie Wang 1,2,* , Liang Wang 1,* , Qian Bu 1, Yuzhou Xiao1, Yue Zhao1, Linhong Jiang1, 5 Yanping Dai 1 , Hongchun Li 1 , Haxiaoyu Liu 1 , Yaxing Chen 1 , Angelo D. Flores 2 , Yinglan Zhao 1 , 6 Xiaobo Cen 1,# 7 1.

    Pull Down Assay:

    Article Title: LUZP1 Regulates Dendritic Spine Maturation and Synaptic Plasticity in the Hippocampal Dentate Gyrus of Mice
    Article Snippet: Research Articles | Development/Plasticity/Repair LUZP1 Regulates Dendritic Spine Maturation and Synaptic Plasticity in the Hippocampal Dentate Gyrus of Mice https://doi.org/10.1523/JNEUROSCI.1867-24.2025 Received: 30 September 2024 Revised: 16 February 2025 Accepted: 18 March 2025 Copyright © 2025 the authors This Early Release article has been peer reviewed and accepted, but has not been through the composition and copyediting processes.The final version may differ slightly in style or formatting and will contain links to any extended data.. Alerts: Sign up at www.jneurosci.org/alerts to receive customized email alerts when the fully formatted version of this article is published.. - 1 - LUZP1 Regulates Dendritic Spine Maturation and 1 Synaptic Plasticity in the Hippocampal Dentate Gyrus of 2 Mice 3 Abbreviated title: LUZP1 in Hippocampal Synaptic Plasticity 4 Xiaojie Wang 1,2,* , Liang Wang 1,* , Qian Bu 1, Yuzhou Xiao1, Yue Zhao1, Linhong Jiang1, 5 Yanping Dai 1 , Hongchun Li 1 , Haxiaoyu Liu 1 , Yaxing Chen 1 , Angelo D. Flores 2 , Yinglan Zhao 1 , 6 Xiaobo Cen 1,# 7 1.



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    a , Immunoblots showing cleaved Notch1 (c-Notch1) and total Notch1 in MVECs treated with S1P for 1-hour ± γ-secretase inhibitor DAPT; quantification below (mean ± SEM). b , Fluorescent micrographs of YFP-Notch1 reporter intensity after S1P treatment; quantification below. Scale bar, 50 μm. c-d , Fold change in HES1 and HEY1 gene expression levels in (c) MVECs after S1P ± DAPT treatment, and in (d) mouse tissues after intravenous injection of S1P ± DAPT. e , Evans blue dye leakage in mouse dermis 1-hour post-injection with DMSO, S1P, or S1P + DAPT. Scale bar, 5 mm. f-i , Quantified dye leakage in (f) dermis, (g) lung, (h) liver, and (i) kidney, measured via absorbance. j , Heat maps of dextran dye diffusion in engineered microvessels ± S1P. Scale bar, 50 μm. k-l , Diffusive permeability in microvessels lined with (k) DAPT-vs. DMSO-treated or (l) NOTCH1 KO vs. SCR KO MVECs (mean ± SD). m , Micrographs of VE-cadherin (magenta), actin (green), and nuclei (blue) staining in microvessels. Scale bar, 50 μm. n-o , Mean corrected intensities of junctional (n) VE-cadherin and (o) actin from (m). p , <t>Rac1</t> activity (PBD pull-down). Active/total Rac1 quantification below. q , Schematic of canonical Notch signaling. r , HES1 / HEY1 gene expression levels in dominant negative-MAML-GFP (DN-MAML-GFP) normalized to GFP expressing cells. s , Permeability in DN-MAML-GFP vs GFP-lined microvessels ± S1P. t , Schematic of Notch cortical pathway mediated by its transmembrane domain (TMD). u , Permeability in TMD-RFP vs RFP-lined microvessels ± S1P. v , Permeability in TMD-RFP vs RFP-lined microvessels ± clinical S1P receptor degrader fingolimod.
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    a , Immunoblots showing cleaved Notch1 (c-Notch1) and total Notch1 in MVECs treated with S1P for 1-hour ± γ-secretase inhibitor DAPT; quantification below (mean ± SEM). b , Fluorescent micrographs of YFP-Notch1 reporter intensity after S1P treatment; quantification below. Scale bar, 50 μm. c-d , Fold change in HES1 and HEY1 gene expression levels in (c) MVECs after S1P ± DAPT treatment, and in (d) mouse tissues after intravenous injection of S1P ± DAPT. e , Evans blue dye leakage in mouse dermis 1-hour post-injection with DMSO, S1P, or S1P + DAPT. Scale bar, 5 mm. f-i , Quantified dye leakage in (f) dermis, (g) lung, (h) liver, and (i) kidney, measured via absorbance. j , Heat maps of dextran dye diffusion in engineered microvessels ± S1P. Scale bar, 50 μm. k-l , Diffusive permeability in microvessels lined with (k) DAPT-vs. DMSO-treated or (l) NOTCH1 KO vs. SCR KO MVECs (mean ± SD). m , Micrographs of VE-cadherin (magenta), actin (green), and nuclei (blue) staining in microvessels. Scale bar, 50 μm. n-o , Mean corrected intensities of junctional (n) VE-cadherin and (o) actin from (m). p , <t>Rac1</t> activity (PBD pull-down). Active/total Rac1 quantification below. q , Schematic of canonical Notch signaling. r , HES1 / HEY1 gene expression levels in dominant negative-MAML-GFP (DN-MAML-GFP) normalized to GFP expressing cells. s , Permeability in DN-MAML-GFP vs GFP-lined microvessels ± S1P. t , Schematic of Notch cortical pathway mediated by its transmembrane domain (TMD). u , Permeability in TMD-RFP vs RFP-lined microvessels ± S1P. v , Permeability in TMD-RFP vs RFP-lined microvessels ± clinical S1P receptor degrader fingolimod.
    Pak Pbd Protein Gst Beads Pak02, supplied by Cytoskeleton Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    a , Immunoblots showing cleaved Notch1 (c-Notch1) and total Notch1 in MVECs treated with S1P for 1-hour ± γ-secretase inhibitor DAPT; quantification below (mean ± SEM). b , Fluorescent micrographs of YFP-Notch1 reporter intensity after S1P treatment; quantification below. Scale bar, 50 μm. c-d , Fold change in HES1 and HEY1 gene expression levels in (c) MVECs after S1P ± DAPT treatment, and in (d) mouse tissues after intravenous injection of S1P ± DAPT. e , Evans blue dye leakage in mouse dermis 1-hour post-injection with DMSO, S1P, or S1P + DAPT. Scale bar, 5 mm. f-i , Quantified dye leakage in (f) dermis, (g) lung, (h) liver, and (i) kidney, measured via absorbance. j , Heat maps of dextran dye diffusion in engineered microvessels ± S1P. Scale bar, 50 μm. k-l , Diffusive permeability in microvessels lined with (k) DAPT-vs. DMSO-treated or (l) NOTCH1 KO vs. SCR KO MVECs (mean ± SD). m , Micrographs of VE-cadherin (magenta), actin (green), and nuclei (blue) staining in microvessels. Scale bar, 50 μm. n-o , Mean corrected intensities of junctional (n) VE-cadherin and (o) actin from (m). p , <t>Rac1</t> activity (PBD pull-down). Active/total Rac1 quantification below. q , Schematic of canonical Notch signaling. r , HES1 / HEY1 gene expression levels in dominant negative-MAML-GFP (DN-MAML-GFP) normalized to GFP expressing cells. s , Permeability in DN-MAML-GFP vs GFP-lined microvessels ± S1P. t , Schematic of Notch cortical pathway mediated by its transmembrane domain (TMD). u , Permeability in TMD-RFP vs RFP-lined microvessels ± S1P. v , Permeability in TMD-RFP vs RFP-lined microvessels ± clinical S1P receptor degrader fingolimod.
    Human Pak1, supplied by Cytoskeleton Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pak+pbd+beads/PAK-PBD+beads+-+binds+active+Rac+Cdc42+proteins/pm41174051-280-19-24
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    a , Immunoblots showing cleaved Notch1 (c-Notch1) and total Notch1 in MVECs treated with S1P for 1-hour ± γ-secretase inhibitor DAPT; quantification below (mean ± SEM). b , Fluorescent micrographs of YFP-Notch1 reporter intensity after S1P treatment; quantification below. Scale bar, 50 μm. c-d , Fold change in HES1 and HEY1 gene expression levels in (c) MVECs after S1P ± DAPT treatment, and in (d) mouse tissues after intravenous injection of S1P ± DAPT. e , Evans blue dye leakage in mouse dermis 1-hour post-injection with DMSO, S1P, or S1P + DAPT. Scale bar, 5 mm. f-i , Quantified dye leakage in (f) dermis, (g) lung, (h) liver, and (i) kidney, measured via absorbance. j , Heat maps of dextran dye diffusion in engineered microvessels ± S1P. Scale bar, 50 μm. k-l , Diffusive permeability in microvessels lined with (k) DAPT-vs. DMSO-treated or (l) NOTCH1 KO vs. SCR KO MVECs (mean ± SD). m , Micrographs of VE-cadherin (magenta), actin (green), and nuclei (blue) staining in microvessels. Scale bar, 50 μm. n-o , Mean corrected intensities of junctional (n) VE-cadherin and (o) actin from (m). p , Rac1 activity (PBD pull-down). Active/total Rac1 quantification below. q , Schematic of canonical Notch signaling. r , HES1 / HEY1 gene expression levels in dominant negative-MAML-GFP (DN-MAML-GFP) normalized to GFP expressing cells. s , Permeability in DN-MAML-GFP vs GFP-lined microvessels ± S1P. t , Schematic of Notch cortical pathway mediated by its transmembrane domain (TMD). u , Permeability in TMD-RFP vs RFP-lined microvessels ± S1P. v , Permeability in TMD-RFP vs RFP-lined microvessels ± clinical S1P receptor degrader fingolimod.

    Journal: bioRxiv

    Article Title: Sphingosine-1-phosphate cross-talks to Notch via a S1PR1-Dll4-MPDZ complex to regulate endothelial barrier function

    doi: 10.64898/2026.05.20.726610

    Figure Lengend Snippet: a , Immunoblots showing cleaved Notch1 (c-Notch1) and total Notch1 in MVECs treated with S1P for 1-hour ± γ-secretase inhibitor DAPT; quantification below (mean ± SEM). b , Fluorescent micrographs of YFP-Notch1 reporter intensity after S1P treatment; quantification below. Scale bar, 50 μm. c-d , Fold change in HES1 and HEY1 gene expression levels in (c) MVECs after S1P ± DAPT treatment, and in (d) mouse tissues after intravenous injection of S1P ± DAPT. e , Evans blue dye leakage in mouse dermis 1-hour post-injection with DMSO, S1P, or S1P + DAPT. Scale bar, 5 mm. f-i , Quantified dye leakage in (f) dermis, (g) lung, (h) liver, and (i) kidney, measured via absorbance. j , Heat maps of dextran dye diffusion in engineered microvessels ± S1P. Scale bar, 50 μm. k-l , Diffusive permeability in microvessels lined with (k) DAPT-vs. DMSO-treated or (l) NOTCH1 KO vs. SCR KO MVECs (mean ± SD). m , Micrographs of VE-cadherin (magenta), actin (green), and nuclei (blue) staining in microvessels. Scale bar, 50 μm. n-o , Mean corrected intensities of junctional (n) VE-cadherin and (o) actin from (m). p , Rac1 activity (PBD pull-down). Active/total Rac1 quantification below. q , Schematic of canonical Notch signaling. r , HES1 / HEY1 gene expression levels in dominant negative-MAML-GFP (DN-MAML-GFP) normalized to GFP expressing cells. s , Permeability in DN-MAML-GFP vs GFP-lined microvessels ± S1P. t , Schematic of Notch cortical pathway mediated by its transmembrane domain (TMD). u , Permeability in TMD-RFP vs RFP-lined microvessels ± S1P. v , Permeability in TMD-RFP vs RFP-lined microvessels ± clinical S1P receptor degrader fingolimod.

    Article Snippet: PAK-PBD beads to bind active Rac1 (PAK-02) were purchased from Cytoskeleton and reconstituted according to the manufacturer’s instructions.

    Techniques: Western Blot, Gene Expression, Injection, Diffusion-based Assay, Permeability, Staining, Activity Assay, Dominant Negative Mutation, Expressing