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phosphorylated pkcz  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phosphorylated pkcz
    Fig. 4. CCM1 is required for junctional localization of the components of the polarity complex Tiam, Par3 and <t>phosphorylated</t> <t>PKCz</t> in cultured ECs and in human CCM1 vascular lesion. (A,B) Junctional staining of Tiam, Par3 and phosphorylated PKCz, in both Ccm1-silenced VEC-positive cells and in Ccm1 fl/fl+Cre cells, appears convoluted and fragmented (arrowheads). This pattern is reminiscent of VEC and b-catenin distribution in Ccm1-silenced ECs (Fig. 3D,E). Scale bar: 20 mm. (C) Phosphorylation of PKCz at Thr410 (p-PKCz) is downregulated by 40% and 80%, respectively, in CDH5-silenced HUVECs and in VEC-null cells. Phosphorylation of PKCz is not modified after CCM1 or Ccm1 siRNA treatment. Data shown are representative of three independent experiments. Vertical bars indicate that the bands, from the same blot, do not come from contiguous lanes. (D) Brain vascular lesion of a patient with heterozygous mutation of CCM1 gene. Phosphorylated PKCz expression is diffuse over large areas in the endothelium of the vascular mulberry lesion (arrowheads, magnification of boxed area). It is instead concentrated to interendothelial junctions in peri-lesion vessels (arrowheads, magnification of boxed area). Scale bars: 50 mm and 10 mm (in magnifications). Asterisks indicate the nuclei of ECs lining the lumen (dashed line).
    Phosphorylated Pkcz, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 128 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "CCM1 regulates vascular-lumen organization by inducing endothelial polarity."

    Article Title: CCM1 regulates vascular-lumen organization by inducing endothelial polarity.

    Journal: Journal of cell science

    doi: 10.1242/jcs.059329

    Fig. 4. CCM1 is required for junctional localization of the components of the polarity complex Tiam, Par3 and phosphorylated PKCz in cultured ECs and in human CCM1 vascular lesion. (A,B) Junctional staining of Tiam, Par3 and phosphorylated PKCz, in both Ccm1-silenced VEC-positive cells and in Ccm1 fl/fl+Cre cells, appears convoluted and fragmented (arrowheads). This pattern is reminiscent of VEC and b-catenin distribution in Ccm1-silenced ECs (Fig. 3D,E). Scale bar: 20 mm. (C) Phosphorylation of PKCz at Thr410 (p-PKCz) is downregulated by 40% and 80%, respectively, in CDH5-silenced HUVECs and in VEC-null cells. Phosphorylation of PKCz is not modified after CCM1 or Ccm1 siRNA treatment. Data shown are representative of three independent experiments. Vertical bars indicate that the bands, from the same blot, do not come from contiguous lanes. (D) Brain vascular lesion of a patient with heterozygous mutation of CCM1 gene. Phosphorylated PKCz expression is diffuse over large areas in the endothelium of the vascular mulberry lesion (arrowheads, magnification of boxed area). It is instead concentrated to interendothelial junctions in peri-lesion vessels (arrowheads, magnification of boxed area). Scale bars: 50 mm and 10 mm (in magnifications). Asterisks indicate the nuclei of ECs lining the lumen (dashed line).
    Figure Legend Snippet: Fig. 4. CCM1 is required for junctional localization of the components of the polarity complex Tiam, Par3 and phosphorylated PKCz in cultured ECs and in human CCM1 vascular lesion. (A,B) Junctional staining of Tiam, Par3 and phosphorylated PKCz, in both Ccm1-silenced VEC-positive cells and in Ccm1 fl/fl+Cre cells, appears convoluted and fragmented (arrowheads). This pattern is reminiscent of VEC and b-catenin distribution in Ccm1-silenced ECs (Fig. 3D,E). Scale bar: 20 mm. (C) Phosphorylation of PKCz at Thr410 (p-PKCz) is downregulated by 40% and 80%, respectively, in CDH5-silenced HUVECs and in VEC-null cells. Phosphorylation of PKCz is not modified after CCM1 or Ccm1 siRNA treatment. Data shown are representative of three independent experiments. Vertical bars indicate that the bands, from the same blot, do not come from contiguous lanes. (D) Brain vascular lesion of a patient with heterozygous mutation of CCM1 gene. Phosphorylated PKCz expression is diffuse over large areas in the endothelium of the vascular mulberry lesion (arrowheads, magnification of boxed area). It is instead concentrated to interendothelial junctions in peri-lesion vessels (arrowheads, magnification of boxed area). Scale bars: 50 mm and 10 mm (in magnifications). Asterisks indicate the nuclei of ECs lining the lumen (dashed line).

    Techniques Used: Cell Culture, Staining, Phospho-proteomics, Modification, Mutagenesis, Expressing

    Fig. 5. Rap1b contributes to the polarizing activity of VEC and CCM1. (A) Rap1b regulates the junctional localization of phosphorylated PKCz. In RAP1B-silenced HUVECs, phosphorylated PKCz loses its linear junctional expression pattern (arrowheads), which instead becomes fragmented (arrows). Scale bar: 20 mm. (B) Rap1b requires VEC expression for its activation. Pull-down of GTP-Rap1was followed by western blotting for Rap1b. Two additional experiments gave comparable results. (C) RAP1B-silenced HUVECs show abnormal lumen (L) maturation (24 hours). Podxl and Coll IV apical or basal localization, respectively, is lost (arrowheads and arrows). The dashed lines outline the lumen. Scale bars: 50 mm and 10 mm (magnifications). (D) Rap1 disappears from cell-cell contacts in Ccm1 silenced or Ccm1 fl/fl+Cre cells and often concentrates at focal contacts (arrows and arrowheads). (E) Active Rap1 (HA-Rap1 GV12) concentrates to cell-cell junctions with VEC, in control cells (Ccm1 fl/fl) (arrows). The junctional localization is strongly reduced or lost (arrows) in parallel with diffusion of VEC in Ccm1-silenced (Ccm1 fl/fl+Cre) ECs. Immunofluorescence staining with an antibody against the HA tag present in Rap1 GV12. Scale bar: 20 mm (D,E).
    Figure Legend Snippet: Fig. 5. Rap1b contributes to the polarizing activity of VEC and CCM1. (A) Rap1b regulates the junctional localization of phosphorylated PKCz. In RAP1B-silenced HUVECs, phosphorylated PKCz loses its linear junctional expression pattern (arrowheads), which instead becomes fragmented (arrows). Scale bar: 20 mm. (B) Rap1b requires VEC expression for its activation. Pull-down of GTP-Rap1was followed by western blotting for Rap1b. Two additional experiments gave comparable results. (C) RAP1B-silenced HUVECs show abnormal lumen (L) maturation (24 hours). Podxl and Coll IV apical or basal localization, respectively, is lost (arrowheads and arrows). The dashed lines outline the lumen. Scale bars: 50 mm and 10 mm (magnifications). (D) Rap1 disappears from cell-cell contacts in Ccm1 silenced or Ccm1 fl/fl+Cre cells and often concentrates at focal contacts (arrows and arrowheads). (E) Active Rap1 (HA-Rap1 GV12) concentrates to cell-cell junctions with VEC, in control cells (Ccm1 fl/fl) (arrows). The junctional localization is strongly reduced or lost (arrows) in parallel with diffusion of VEC in Ccm1-silenced (Ccm1 fl/fl+Cre) ECs. Immunofluorescence staining with an antibody against the HA tag present in Rap1 GV12. Scale bar: 20 mm (D,E).

    Techniques Used: Activity Assay, Expressing, Activation Assay, Western Blot, Control, Diffusion-based Assay, Immunofluorescence, Staining

    Related Articles

    Immunofluorescence:

    Article Title: CCM1 regulates vascular-lumen organization by inducing endothelial polarity.
    Article Snippet: .. For immunofluorescence (IF), western blot (WB) and immunoprecipitation, the following antibodies were used: VE-cadherin (C-19, sc-6458, goat, Santa Cruz), bcatenin (mouse, BD Transduction Laboratories), CCM1 (residues 1-207 of the human sequence and crossreacting with mouse protein, rabbit, gifts from Elisabeth TournierLasserve, Université de Médicine Paris Diderot, Paris, France and Francesco Retta, University of Torino, Torino, Italy) and (KRIT1, K-16, sc-23997, goat, Santa Cruz), phosphorylated PKCz (rabbit, Cell Signaling), total a-PKCz (C-20, sc-216, rabbit, Santa Cruz), Rap1 (121, sc-65, rabbit, Santa Cruz, for IF), Rap1b (rabbit, Cell Signaling, for WB), Tiam 1 (C-16, sc-872, rabbit, Santa Cruz), Podxl (murine-specific and human-specific, R&D), Coll IV (murine-specific and human-specific, AbD Serotec), Par3 (rabbit, Millipore), Pecam (clone 13.3, rat anti-mouse), Flag, (M2, mouse, Sigma), rac (mouse, BD Transduction Laboratories), cdc42 (rabbit, Cell Signaling), HA (clone 11, mouse, Covance). ..

    Western Blot:

    Article Title: CCM1 regulates vascular-lumen organization by inducing endothelial polarity.
    Article Snippet: .. For immunofluorescence (IF), western blot (WB) and immunoprecipitation, the following antibodies were used: VE-cadherin (C-19, sc-6458, goat, Santa Cruz), bcatenin (mouse, BD Transduction Laboratories), CCM1 (residues 1-207 of the human sequence and crossreacting with mouse protein, rabbit, gifts from Elisabeth TournierLasserve, Université de Médicine Paris Diderot, Paris, France and Francesco Retta, University of Torino, Torino, Italy) and (KRIT1, K-16, sc-23997, goat, Santa Cruz), phosphorylated PKCz (rabbit, Cell Signaling), total a-PKCz (C-20, sc-216, rabbit, Santa Cruz), Rap1 (121, sc-65, rabbit, Santa Cruz, for IF), Rap1b (rabbit, Cell Signaling, for WB), Tiam 1 (C-16, sc-872, rabbit, Santa Cruz), Podxl (murine-specific and human-specific, R&D), Coll IV (murine-specific and human-specific, AbD Serotec), Par3 (rabbit, Millipore), Pecam (clone 13.3, rat anti-mouse), Flag, (M2, mouse, Sigma), rac (mouse, BD Transduction Laboratories), cdc42 (rabbit, Cell Signaling), HA (clone 11, mouse, Covance). ..

    Immunoprecipitation:

    Article Title: CCM1 regulates vascular-lumen organization by inducing endothelial polarity.
    Article Snippet: .. For immunofluorescence (IF), western blot (WB) and immunoprecipitation, the following antibodies were used: VE-cadherin (C-19, sc-6458, goat, Santa Cruz), bcatenin (mouse, BD Transduction Laboratories), CCM1 (residues 1-207 of the human sequence and crossreacting with mouse protein, rabbit, gifts from Elisabeth TournierLasserve, Université de Médicine Paris Diderot, Paris, France and Francesco Retta, University of Torino, Torino, Italy) and (KRIT1, K-16, sc-23997, goat, Santa Cruz), phosphorylated PKCz (rabbit, Cell Signaling), total a-PKCz (C-20, sc-216, rabbit, Santa Cruz), Rap1 (121, sc-65, rabbit, Santa Cruz, for IF), Rap1b (rabbit, Cell Signaling, for WB), Tiam 1 (C-16, sc-872, rabbit, Santa Cruz), Podxl (murine-specific and human-specific, R&D), Coll IV (murine-specific and human-specific, AbD Serotec), Par3 (rabbit, Millipore), Pecam (clone 13.3, rat anti-mouse), Flag, (M2, mouse, Sigma), rac (mouse, BD Transduction Laboratories), cdc42 (rabbit, Cell Signaling), HA (clone 11, mouse, Covance). ..

    Sequencing:

    Article Title: CCM1 regulates vascular-lumen organization by inducing endothelial polarity.
    Article Snippet: .. For immunofluorescence (IF), western blot (WB) and immunoprecipitation, the following antibodies were used: VE-cadherin (C-19, sc-6458, goat, Santa Cruz), bcatenin (mouse, BD Transduction Laboratories), CCM1 (residues 1-207 of the human sequence and crossreacting with mouse protein, rabbit, gifts from Elisabeth TournierLasserve, Université de Médicine Paris Diderot, Paris, France and Francesco Retta, University of Torino, Torino, Italy) and (KRIT1, K-16, sc-23997, goat, Santa Cruz), phosphorylated PKCz (rabbit, Cell Signaling), total a-PKCz (C-20, sc-216, rabbit, Santa Cruz), Rap1 (121, sc-65, rabbit, Santa Cruz, for IF), Rap1b (rabbit, Cell Signaling, for WB), Tiam 1 (C-16, sc-872, rabbit, Santa Cruz), Podxl (murine-specific and human-specific, R&D), Coll IV (murine-specific and human-specific, AbD Serotec), Par3 (rabbit, Millipore), Pecam (clone 13.3, rat anti-mouse), Flag, (M2, mouse, Sigma), rac (mouse, BD Transduction Laboratories), cdc42 (rabbit, Cell Signaling), HA (clone 11, mouse, Covance). ..



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    Fig. 4. CCM1 is required for junctional localization of the components of the polarity complex Tiam, Par3 and <t>phosphorylated</t> <t>PKCz</t> in cultured ECs and in human CCM1 vascular lesion. (A,B) Junctional staining of Tiam, Par3 and phosphorylated PKCz, in both Ccm1-silenced VEC-positive cells and in Ccm1 fl/fl+Cre cells, appears convoluted and fragmented (arrowheads). This pattern is reminiscent of VEC and b-catenin distribution in Ccm1-silenced ECs (Fig. 3D,E). Scale bar: 20 mm. (C) Phosphorylation of PKCz at Thr410 (p-PKCz) is downregulated by 40% and 80%, respectively, in CDH5-silenced HUVECs and in VEC-null cells. Phosphorylation of PKCz is not modified after CCM1 or Ccm1 siRNA treatment. Data shown are representative of three independent experiments. Vertical bars indicate that the bands, from the same blot, do not come from contiguous lanes. (D) Brain vascular lesion of a patient with heterozygous mutation of CCM1 gene. Phosphorylated PKCz expression is diffuse over large areas in the endothelium of the vascular mulberry lesion (arrowheads, magnification of boxed area). It is instead concentrated to interendothelial junctions in peri-lesion vessels (arrowheads, magnification of boxed area). Scale bars: 50 mm and 10 mm (in magnifications). Asterisks indicate the nuclei of ECs lining the lumen (dashed line).
    Phosphorylated Pkcz, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/phosphorylated+pkcz/Phospho-PKCzeta%2Flamba+(Thr410%2F403)+Antibody/pm20332120-165-63-66
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    phosphorylated pkcz - by Bioz Stars, 2026-11
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    Image Search Results


    Fig. 4. CCM1 is required for junctional localization of the components of the polarity complex Tiam, Par3 and phosphorylated PKCz in cultured ECs and in human CCM1 vascular lesion. (A,B) Junctional staining of Tiam, Par3 and phosphorylated PKCz, in both Ccm1-silenced VEC-positive cells and in Ccm1 fl/fl+Cre cells, appears convoluted and fragmented (arrowheads). This pattern is reminiscent of VEC and b-catenin distribution in Ccm1-silenced ECs (Fig. 3D,E). Scale bar: 20 mm. (C) Phosphorylation of PKCz at Thr410 (p-PKCz) is downregulated by 40% and 80%, respectively, in CDH5-silenced HUVECs and in VEC-null cells. Phosphorylation of PKCz is not modified after CCM1 or Ccm1 siRNA treatment. Data shown are representative of three independent experiments. Vertical bars indicate that the bands, from the same blot, do not come from contiguous lanes. (D) Brain vascular lesion of a patient with heterozygous mutation of CCM1 gene. Phosphorylated PKCz expression is diffuse over large areas in the endothelium of the vascular mulberry lesion (arrowheads, magnification of boxed area). It is instead concentrated to interendothelial junctions in peri-lesion vessels (arrowheads, magnification of boxed area). Scale bars: 50 mm and 10 mm (in magnifications). Asterisks indicate the nuclei of ECs lining the lumen (dashed line).

    Journal: Journal of cell science

    Article Title: CCM1 regulates vascular-lumen organization by inducing endothelial polarity.

    doi: 10.1242/jcs.059329

    Figure Lengend Snippet: Fig. 4. CCM1 is required for junctional localization of the components of the polarity complex Tiam, Par3 and phosphorylated PKCz in cultured ECs and in human CCM1 vascular lesion. (A,B) Junctional staining of Tiam, Par3 and phosphorylated PKCz, in both Ccm1-silenced VEC-positive cells and in Ccm1 fl/fl+Cre cells, appears convoluted and fragmented (arrowheads). This pattern is reminiscent of VEC and b-catenin distribution in Ccm1-silenced ECs (Fig. 3D,E). Scale bar: 20 mm. (C) Phosphorylation of PKCz at Thr410 (p-PKCz) is downregulated by 40% and 80%, respectively, in CDH5-silenced HUVECs and in VEC-null cells. Phosphorylation of PKCz is not modified after CCM1 or Ccm1 siRNA treatment. Data shown are representative of three independent experiments. Vertical bars indicate that the bands, from the same blot, do not come from contiguous lanes. (D) Brain vascular lesion of a patient with heterozygous mutation of CCM1 gene. Phosphorylated PKCz expression is diffuse over large areas in the endothelium of the vascular mulberry lesion (arrowheads, magnification of boxed area). It is instead concentrated to interendothelial junctions in peri-lesion vessels (arrowheads, magnification of boxed area). Scale bars: 50 mm and 10 mm (in magnifications). Asterisks indicate the nuclei of ECs lining the lumen (dashed line).

    Article Snippet: For immunofluorescence (IF), western blot (WB) and immunoprecipitation, the following antibodies were used: VE-cadherin (C-19, sc-6458, goat, Santa Cruz), bcatenin (mouse, BD Transduction Laboratories), CCM1 (residues 1-207 of the human sequence and crossreacting with mouse protein, rabbit, gifts from Elisabeth TournierLasserve, Université de Médicine Paris Diderot, Paris, France and Francesco Retta, University of Torino, Torino, Italy) and (KRIT1, K-16, sc-23997, goat, Santa Cruz), phosphorylated PKCz (rabbit, Cell Signaling), total a-PKCz (C-20, sc-216, rabbit, Santa Cruz), Rap1 (121, sc-65, rabbit, Santa Cruz, for IF), Rap1b (rabbit, Cell Signaling, for WB), Tiam 1 (C-16, sc-872, rabbit, Santa Cruz), Podxl (murine-specific and human-specific, R&D), Coll IV (murine-specific and human-specific, AbD Serotec), Par3 (rabbit, Millipore), Pecam (clone 13.3, rat anti-mouse), Flag, (M2, mouse, Sigma), rac (mouse, BD Transduction Laboratories), cdc42 (rabbit, Cell Signaling), HA (clone 11, mouse, Covance).

    Techniques: Cell Culture, Staining, Phospho-proteomics, Modification, Mutagenesis, Expressing

    Fig. 5. Rap1b contributes to the polarizing activity of VEC and CCM1. (A) Rap1b regulates the junctional localization of phosphorylated PKCz. In RAP1B-silenced HUVECs, phosphorylated PKCz loses its linear junctional expression pattern (arrowheads), which instead becomes fragmented (arrows). Scale bar: 20 mm. (B) Rap1b requires VEC expression for its activation. Pull-down of GTP-Rap1was followed by western blotting for Rap1b. Two additional experiments gave comparable results. (C) RAP1B-silenced HUVECs show abnormal lumen (L) maturation (24 hours). Podxl and Coll IV apical or basal localization, respectively, is lost (arrowheads and arrows). The dashed lines outline the lumen. Scale bars: 50 mm and 10 mm (magnifications). (D) Rap1 disappears from cell-cell contacts in Ccm1 silenced or Ccm1 fl/fl+Cre cells and often concentrates at focal contacts (arrows and arrowheads). (E) Active Rap1 (HA-Rap1 GV12) concentrates to cell-cell junctions with VEC, in control cells (Ccm1 fl/fl) (arrows). The junctional localization is strongly reduced or lost (arrows) in parallel with diffusion of VEC in Ccm1-silenced (Ccm1 fl/fl+Cre) ECs. Immunofluorescence staining with an antibody against the HA tag present in Rap1 GV12. Scale bar: 20 mm (D,E).

    Journal: Journal of cell science

    Article Title: CCM1 regulates vascular-lumen organization by inducing endothelial polarity.

    doi: 10.1242/jcs.059329

    Figure Lengend Snippet: Fig. 5. Rap1b contributes to the polarizing activity of VEC and CCM1. (A) Rap1b regulates the junctional localization of phosphorylated PKCz. In RAP1B-silenced HUVECs, phosphorylated PKCz loses its linear junctional expression pattern (arrowheads), which instead becomes fragmented (arrows). Scale bar: 20 mm. (B) Rap1b requires VEC expression for its activation. Pull-down of GTP-Rap1was followed by western blotting for Rap1b. Two additional experiments gave comparable results. (C) RAP1B-silenced HUVECs show abnormal lumen (L) maturation (24 hours). Podxl and Coll IV apical or basal localization, respectively, is lost (arrowheads and arrows). The dashed lines outline the lumen. Scale bars: 50 mm and 10 mm (magnifications). (D) Rap1 disappears from cell-cell contacts in Ccm1 silenced or Ccm1 fl/fl+Cre cells and often concentrates at focal contacts (arrows and arrowheads). (E) Active Rap1 (HA-Rap1 GV12) concentrates to cell-cell junctions with VEC, in control cells (Ccm1 fl/fl) (arrows). The junctional localization is strongly reduced or lost (arrows) in parallel with diffusion of VEC in Ccm1-silenced (Ccm1 fl/fl+Cre) ECs. Immunofluorescence staining with an antibody against the HA tag present in Rap1 GV12. Scale bar: 20 mm (D,E).

    Article Snippet: For immunofluorescence (IF), western blot (WB) and immunoprecipitation, the following antibodies were used: VE-cadherin (C-19, sc-6458, goat, Santa Cruz), bcatenin (mouse, BD Transduction Laboratories), CCM1 (residues 1-207 of the human sequence and crossreacting with mouse protein, rabbit, gifts from Elisabeth TournierLasserve, Université de Médicine Paris Diderot, Paris, France and Francesco Retta, University of Torino, Torino, Italy) and (KRIT1, K-16, sc-23997, goat, Santa Cruz), phosphorylated PKCz (rabbit, Cell Signaling), total a-PKCz (C-20, sc-216, rabbit, Santa Cruz), Rap1 (121, sc-65, rabbit, Santa Cruz, for IF), Rap1b (rabbit, Cell Signaling, for WB), Tiam 1 (C-16, sc-872, rabbit, Santa Cruz), Podxl (murine-specific and human-specific, R&D), Coll IV (murine-specific and human-specific, AbD Serotec), Par3 (rabbit, Millipore), Pecam (clone 13.3, rat anti-mouse), Flag, (M2, mouse, Sigma), rac (mouse, BD Transduction Laboratories), cdc42 (rabbit, Cell Signaling), HA (clone 11, mouse, Covance).

    Techniques: Activity Assay, Expressing, Activation Assay, Western Blot, Control, Diffusion-based Assay, Immunofluorescence, Staining