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cn03  (Cytoskeleton Inc)


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    Structured Review

    Cytoskeleton Inc cn03
    Cell-substrate adhesion characteristics control cell spanning. (a) Representative microscopy images for fibroblasts adhered to a soft substrate, stimulated with <t>CN03</t> (RhoA activator), Y27632 (ROCK inhibitor), PF228 (Focal adhesion kinase inhibitor), and MMC (DNA crosslinking). (b) Cell length for the distinct modulators and (c) FSD compared to the fibroblasts (dash-dotted reference line indicates the median value). (d) Relationship between probability of spanning cells and cylinder diameter for the selected cell types. (e) Correlation between cell length and probability of cells spanning. (f) Correlation between FSD and probability of cells spanning. In red, allometric fit, given by y = a x b . (g, left) Schematic representation of the boundary conditions ( U, UR indicating translational and rotational degrees of freedom respectively) and material properties of the FE model representing an individual cell with two adhesion morphologies (C1 = fully adherent, circular morphology, C2 = large FSD, polar morphology) attached to cylindrical surfaces with Ø = 100 and 1000 μm. (g, middle) Resulting cell displacement according to the cell adhesion morphology (C1, C2) on a cylinder with Ø = 100 μm and (g, right) on a cylinder with Ø = 1000 μm in isometric view (top row) and front view (bottom row). Vector plot is combined with deformed shape to better visualize the direction and magnitude of the displacement. Statistical significance via Mann-Whitney test (two sided) with Bonferroni correction, ∗p < 0.05. N ≥ 60 cells/cell type for FA and morphological analysis. N ≥ 3 GeoChips/cell type for a total of N ≥ 12 half-cylinders/condition. 1 donor/cell type. Scale bar 50 μm.
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    Images

    1) Product Images from "Cell type-specific response to curvature controls tissue growth dynamics in biomaterial pores"

    Article Title: Cell type-specific response to curvature controls tissue growth dynamics in biomaterial pores

    Journal: Bioactive Materials

    doi: 10.1016/j.bioactmat.2026.02.005

    Cell-substrate adhesion characteristics control cell spanning. (a) Representative microscopy images for fibroblasts adhered to a soft substrate, stimulated with CN03 (RhoA activator), Y27632 (ROCK inhibitor), PF228 (Focal adhesion kinase inhibitor), and MMC (DNA crosslinking). (b) Cell length for the distinct modulators and (c) FSD compared to the fibroblasts (dash-dotted reference line indicates the median value). (d) Relationship between probability of spanning cells and cylinder diameter for the selected cell types. (e) Correlation between cell length and probability of cells spanning. (f) Correlation between FSD and probability of cells spanning. In red, allometric fit, given by y = a x b . (g, left) Schematic representation of the boundary conditions ( U, UR indicating translational and rotational degrees of freedom respectively) and material properties of the FE model representing an individual cell with two adhesion morphologies (C1 = fully adherent, circular morphology, C2 = large FSD, polar morphology) attached to cylindrical surfaces with Ø = 100 and 1000 μm. (g, middle) Resulting cell displacement according to the cell adhesion morphology (C1, C2) on a cylinder with Ø = 100 μm and (g, right) on a cylinder with Ø = 1000 μm in isometric view (top row) and front view (bottom row). Vector plot is combined with deformed shape to better visualize the direction and magnitude of the displacement. Statistical significance via Mann-Whitney test (two sided) with Bonferroni correction, ∗p < 0.05. N ≥ 60 cells/cell type for FA and morphological analysis. N ≥ 3 GeoChips/cell type for a total of N ≥ 12 half-cylinders/condition. 1 donor/cell type. Scale bar 50 μm.
    Figure Legend Snippet: Cell-substrate adhesion characteristics control cell spanning. (a) Representative microscopy images for fibroblasts adhered to a soft substrate, stimulated with CN03 (RhoA activator), Y27632 (ROCK inhibitor), PF228 (Focal adhesion kinase inhibitor), and MMC (DNA crosslinking). (b) Cell length for the distinct modulators and (c) FSD compared to the fibroblasts (dash-dotted reference line indicates the median value). (d) Relationship between probability of spanning cells and cylinder diameter for the selected cell types. (e) Correlation between cell length and probability of cells spanning. (f) Correlation between FSD and probability of cells spanning. In red, allometric fit, given by y = a x b . (g, left) Schematic representation of the boundary conditions ( U, UR indicating translational and rotational degrees of freedom respectively) and material properties of the FE model representing an individual cell with two adhesion morphologies (C1 = fully adherent, circular morphology, C2 = large FSD, polar morphology) attached to cylindrical surfaces with Ø = 100 and 1000 μm. (g, middle) Resulting cell displacement according to the cell adhesion morphology (C1, C2) on a cylinder with Ø = 100 μm and (g, right) on a cylinder with Ø = 1000 μm in isometric view (top row) and front view (bottom row). Vector plot is combined with deformed shape to better visualize the direction and magnitude of the displacement. Statistical significance via Mann-Whitney test (two sided) with Bonferroni correction, ∗p < 0.05. N ≥ 60 cells/cell type for FA and morphological analysis. N ≥ 3 GeoChips/cell type for a total of N ≥ 12 half-cylinders/condition. 1 donor/cell type. Scale bar 50 μm.

    Techniques Used: Control, Microscopy, Plasmid Preparation, MANN-WHITNEY

    Related Articles

    Cell Culture:

    Article Title: P-cadherin dependent adhesions are required for single lumen formation and HGF-mediated cell protrusions during epithelial morphogenesis
    Article Snippet: .. Two days after lumen formation, samples were cultured in serum free medium and the next day, Rho activator (1 μg/ml; CN03, Cytoskeleton, Inc.) was added to CDH3-/- cysts. .. ROCK inhibitor (12.5 μg/ml; 10005583, Cayman Chemical) was added to WT cysts.

    Article Title: P-cadherin-dependent adhesions are required for single lumen formation and HGF-mediated cell protrusions during epithelial morphogenesis
    Article Snippet: .. Two days after lumen formation, samples were cultured in serum free medium and the next day, Rho activator (1 μg/ml; CN03, Cytoskeleton, Inc.) was added to CDH3 -/- cysts. .. ROCK inhibitor (12.5 μg/ml; 10005583, Cayman Chemical) was added to WT cysts.

    Activation Assay:

    Article Title:
    Article Snippet: Kolmogorov-Smirnov test was used to determine 11 statistical significance. f, Cartoon illustrating a proposed model for how 12 mechanosensitive, frequency-modulated, oscillatory activation of RhoA can differentially 13 regulate actin cytoskeleton remodeling, leading to differences in subcellular dynamics. .. 14 15 16 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 SI Appendix, Figure S5 | Mechanosensitive activation of SMAD1/5. a, Luciferase 19 assay performed on wtNSCs treated as indicated for 18 hours: Rho activator (0.075 20 units/ml; Cytoskeleton, Inc.), Cdc42 activator (0.75 units/ml; Cytoskeleton, Inc.). ..

    Article Title: Cortactin is in a complex with VE-cadherin and is required for endothelial adherens junction stability through Rap1/Rac1 activation
    Article Snippet: .. To elevate intracellular cAMP concentration, 5 μM Forskolin (Sigma Aldrich Chemie, #F6886), and 10 μM Rolipram (Sigma Aldrich Chemie, #R6520) were applied for 1 h. To stimulate the activation of Rho family small GTPases cells were treated with 0.25 μg/ml Rho/Rac/Cdc42 Activator I (Cytoskeleton Inc, #CN04-A) for 2 h. Triggering of RhoA was achieved with the Rho activator I, Calpeptin at 1 U/ml (#CN01-A, Cytoskeleton Inc.). .. Inhibition of PKA was achieved using 10 μM Dihydrochloride (H89, Santa Cruz, sc-3537).

    Article Title: Cortactin is in a complex with VE-cadherin and is required for endothelial adherens junction stability through Rap1/Rac1 activation.
    Article Snippet: .. To elevate intracellular cAMP concentration, 5 μM Forskolin (Sigma Aldrich Chemie, #F6886), and 10 μM Rolipram (Sigma Aldrich Chemie, #R6520) were applied for 1 h. To stimulate the activation of Rho family small GTPases cells were treated with 0.25 μg/ml Rho/Rac/Cdc42 Activator I (Cytoskeleton Inc, #CN04-A) for 2 h. Triggering of RhoA was achieved with the Rho activator I, Calpeptin at 1 U/ml (#CN01-A, Cytoskeleton Inc.). .. Inhibition of PKA was achieved using 10 μM Dihydrochloride (H89, Santa Cruz, sc-3537).

    Luciferase:

    Article Title:
    Article Snippet: Kolmogorov-Smirnov test was used to determine 11 statistical significance. f, Cartoon illustrating a proposed model for how 12 mechanosensitive, frequency-modulated, oscillatory activation of RhoA can differentially 13 regulate actin cytoskeleton remodeling, leading to differences in subcellular dynamics. .. 14 15 16 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 SI Appendix, Figure S5 | Mechanosensitive activation of SMAD1/5. a, Luciferase 19 assay performed on wtNSCs treated as indicated for 18 hours: Rho activator (0.075 20 units/ml; Cytoskeleton, Inc.), Cdc42 activator (0.75 units/ml; Cytoskeleton, Inc.). ..

    Immunostaining:

    Article Title:
    Article Snippet: .. ***P < 0.005, **P < 0.01, 10 *P < 0.05. ns, not significant. d, Immunostaining for pSMAD1/5 Ser463/465 (magenta), 11 Phalloidin (green) and DAPI (blue, nuclei) in wtNSCs treated as indicated for 15 minutes: 12 Rho activator (0.075 units/ml; Cytoskeleton, Inc.), Cdc42 activator (0.75 units/ml; 13 Cytoskeleton, Inc.), LDN-193189 (‘LDN’, 100 nM, R&D Systems) or SB-431542 (‘SB’, 10 14 μm, R&D Systems). ..

    Concentration Assay:

    Article Title: Cortactin is in a complex with VE-cadherin and is required for endothelial adherens junction stability through Rap1/Rac1 activation
    Article Snippet: .. To elevate intracellular cAMP concentration, 5 μM Forskolin (Sigma Aldrich Chemie, #F6886), and 10 μM Rolipram (Sigma Aldrich Chemie, #R6520) were applied for 1 h. To stimulate the activation of Rho family small GTPases cells were treated with 0.25 μg/ml Rho/Rac/Cdc42 Activator I (Cytoskeleton Inc, #CN04-A) for 2 h. Triggering of RhoA was achieved with the Rho activator I, Calpeptin at 1 U/ml (#CN01-A, Cytoskeleton Inc.). .. Inhibition of PKA was achieved using 10 μM Dihydrochloride (H89, Santa Cruz, sc-3537).

    Article Title: Cortactin is in a complex with VE-cadherin and is required for endothelial adherens junction stability through Rap1/Rac1 activation.
    Article Snippet: .. To elevate intracellular cAMP concentration, 5 μM Forskolin (Sigma Aldrich Chemie, #F6886), and 10 μM Rolipram (Sigma Aldrich Chemie, #R6520) were applied for 1 h. To stimulate the activation of Rho family small GTPases cells were treated with 0.25 μg/ml Rho/Rac/Cdc42 Activator I (Cytoskeleton Inc, #CN04-A) for 2 h. Triggering of RhoA was achieved with the Rho activator I, Calpeptin at 1 U/ml (#CN01-A, Cytoskeleton Inc.). .. Inhibition of PKA was achieved using 10 μM Dihydrochloride (H89, Santa Cruz, sc-3537).

    Fluorescence:

    Article Title: VE-Cadherin-Actin Regulation Promotes Mechanotransduction and Monolayer Maturation Involving a Tension-Sensitive Intermediate State.
    Article Snippet: A spinning disk confocal microscope (SpDM) (Carl Zeiss, Oberkochen, Germany) equipped with a single BTF unit (MOS Technologies), definite focus, 37◦C heated stage, humidity, and 5% CO2 supply was used for VE-cadherin-EGFP and LifeAct-mCherry fluorescence live cell imaging. .. High temporal resolution fluorescence time-lapse imaging was performed every 10-20 sec on HUVECs subjected to shear stress or treated with ROCK inhibitor Y27632 (10 μM, Sigma-Aldrich, St. Louis, USA) or Rho activator (0.25 U/mL, Cytoskeleton Inc., Denver, USA) using a Zeiss Plan Apochromat 40x/1.4 oil DIC (UV) VIS-IR immersion objective (Carl Zeiss, Oberkochen, Germany). ..

    Imaging:

    Article Title: VE-Cadherin-Actin Regulation Promotes Mechanotransduction and Monolayer Maturation Involving a Tension-Sensitive Intermediate State.
    Article Snippet: A spinning disk confocal microscope (SpDM) (Carl Zeiss, Oberkochen, Germany) equipped with a single BTF unit (MOS Technologies), definite focus, 37◦C heated stage, humidity, and 5% CO2 supply was used for VE-cadherin-EGFP and LifeAct-mCherry fluorescence live cell imaging. .. High temporal resolution fluorescence time-lapse imaging was performed every 10-20 sec on HUVECs subjected to shear stress or treated with ROCK inhibitor Y27632 (10 μM, Sigma-Aldrich, St. Louis, USA) or Rho activator (0.25 U/mL, Cytoskeleton Inc., Denver, USA) using a Zeiss Plan Apochromat 40x/1.4 oil DIC (UV) VIS-IR immersion objective (Carl Zeiss, Oberkochen, Germany). ..

    Shear:

    Article Title: VE-Cadherin-Actin Regulation Promotes Mechanotransduction and Monolayer Maturation Involving a Tension-Sensitive Intermediate State.
    Article Snippet: A spinning disk confocal microscope (SpDM) (Carl Zeiss, Oberkochen, Germany) equipped with a single BTF unit (MOS Technologies), definite focus, 37◦C heated stage, humidity, and 5% CO2 supply was used for VE-cadherin-EGFP and LifeAct-mCherry fluorescence live cell imaging. .. High temporal resolution fluorescence time-lapse imaging was performed every 10-20 sec on HUVECs subjected to shear stress or treated with ROCK inhibitor Y27632 (10 μM, Sigma-Aldrich, St. Louis, USA) or Rho activator (0.25 U/mL, Cytoskeleton Inc., Denver, USA) using a Zeiss Plan Apochromat 40x/1.4 oil DIC (UV) VIS-IR immersion objective (Carl Zeiss, Oberkochen, Germany). ..



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    Cell-substrate adhesion characteristics control cell spanning. (a) Representative microscopy images for fibroblasts adhered to a soft substrate, stimulated with CN03 (RhoA activator), Y27632 (ROCK inhibitor), PF228 (Focal adhesion kinase inhibitor), and MMC (DNA crosslinking). (b) Cell length for the distinct modulators and (c) FSD compared to the fibroblasts (dash-dotted reference line indicates the median value). (d) Relationship between probability of spanning cells and cylinder diameter for the selected cell types. (e) Correlation between cell length and probability of cells spanning. (f) Correlation between FSD and probability of cells spanning. In red, allometric fit, given by y = a x b . (g, left) Schematic representation of the boundary conditions ( U, UR indicating translational and rotational degrees of freedom respectively) and material properties of the FE model representing an individual cell with two adhesion morphologies (C1 = fully adherent, circular morphology, C2 = large FSD, polar morphology) attached to cylindrical surfaces with Ø = 100 and 1000 μm. (g, middle) Resulting cell displacement according to the cell adhesion morphology (C1, C2) on a cylinder with Ø = 100 μm and (g, right) on a cylinder with Ø = 1000 μm in isometric view (top row) and front view (bottom row). Vector plot is combined with deformed shape to better visualize the direction and magnitude of the displacement. Statistical significance via Mann-Whitney test (two sided) with Bonferroni correction, ∗p < 0.05. N ≥ 60 cells/cell type for FA and morphological analysis. N ≥ 3 GeoChips/cell type for a total of N ≥ 12 half-cylinders/condition. 1 donor/cell type. Scale bar 50 μm.

    Journal: Bioactive Materials

    Article Title: Cell type-specific response to curvature controls tissue growth dynamics in biomaterial pores

    doi: 10.1016/j.bioactmat.2026.02.005

    Figure Lengend Snippet: Cell-substrate adhesion characteristics control cell spanning. (a) Representative microscopy images for fibroblasts adhered to a soft substrate, stimulated with CN03 (RhoA activator), Y27632 (ROCK inhibitor), PF228 (Focal adhesion kinase inhibitor), and MMC (DNA crosslinking). (b) Cell length for the distinct modulators and (c) FSD compared to the fibroblasts (dash-dotted reference line indicates the median value). (d) Relationship between probability of spanning cells and cylinder diameter for the selected cell types. (e) Correlation between cell length and probability of cells spanning. (f) Correlation between FSD and probability of cells spanning. In red, allometric fit, given by y = a x b . (g, left) Schematic representation of the boundary conditions ( U, UR indicating translational and rotational degrees of freedom respectively) and material properties of the FE model representing an individual cell with two adhesion morphologies (C1 = fully adherent, circular morphology, C2 = large FSD, polar morphology) attached to cylindrical surfaces with Ø = 100 and 1000 μm. (g, middle) Resulting cell displacement according to the cell adhesion morphology (C1, C2) on a cylinder with Ø = 100 μm and (g, right) on a cylinder with Ø = 1000 μm in isometric view (top row) and front view (bottom row). Vector plot is combined with deformed shape to better visualize the direction and magnitude of the displacement. Statistical significance via Mann-Whitney test (two sided) with Bonferroni correction, ∗p < 0.05. N ≥ 60 cells/cell type for FA and morphological analysis. N ≥ 3 GeoChips/cell type for a total of N ≥ 12 half-cylinders/condition. 1 donor/cell type. Scale bar 50 μm.

    Article Snippet: Y27632 (13624, Cell Signaling Technology, Inc.) was supplemented to the medium at a concentration of 10 μM, CN03 (Rho Activator II, Cytoskeleton, Inc.) was used at a concentration of 5 μg/ml, and PF228 (PZ0117, Sigma-Aldrich) was supplemented at a concentration of 100 μM.

    Techniques: Control, Microscopy, Plasmid Preparation, MANN-WHITNEY