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phosphorylated src y416 antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phosphorylated src y416 antibody
    Phosphorylated Src Y416 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/phosphorylated+src+y416+antibody/anti+phospho+src/us11981684-944-96-99
    Average 90 stars, based on 1 article reviews
    phosphorylated src y416 antibody - by Bioz Stars, 2026-10
    90/100 stars

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    Article Title: Macrocyclic compounds and uses thereof
    Article Snippet: Protein lysates (approximately 20 μg) was resolved on 4-12% Bolt Bis-Tris precasted gels with MES running buffer (Life Technologies), transferred to nitrocellulose membranes using Trans-Blot Turbo Transfer System (Bio-Rad) and detected with antibodies targeting phosphorylated MET (Y1234/Y1235) (Cell Signaling Technology), MET (Y1349), MET (Y1003), total MET (Cell Signaling Technology), phosphorylated EGFR (Y1068) and total EGFR(Cell Signaling Technology), phosphorylated STAT3 and STATS, total STAT3 and STATS (Cell Signaling Technology), phosphorylated AKT (Cell Signaling Technology), total AKT (Cell Signaling Technology), phosphorylated ERK (Cell Signaling Technology), total ERK (Cell Signaling Technology), phosphorylated PLCγ2 and total PLCγ2 (Cell Signaling Technology), phosphorylated SRC Y416 (Cell Signaling Technology), total SRC (Cell Signaling Technology), phosphorylated paxillin Y118 (Cell Signaling Technology), total paxillin (Cell Signaling Technology), PARP, actin (Cell Signaling Technology).

    Article Title: Macrocyclic compounds and uses thereof
    Article Snippet: Protein lysates (approximately 20 μg) was resolved on 4-12% Bolt Bis-Tris precasted gels with MES running buffer (Life Technologies), transferred to nitrocellulose membranes using Trans-Blot Turbo Transfer System (Bio-Rad) and detected with antibodies targeting phosphorylated MET (Y1234/Y1235) (Cell Signaling Technology), MET (Y1349), MET (Y1003), total MET (Cell Signaling Technology), phosphorylated EGFR (Y1068) and total EGFR(Cell Signaling Technology), phosphorylated STAT3 and STATS, total STAT3 and STAT5 (Cell Signaling Technology), phosphorylated AKT (Cell Signaling Technology), total AKT (Cell Signaling Technology), phosphorylated ERK (Cell Signaling Technology), total ERK (Cell Signaling Technology), phosphorylated PLCγ2 and total PLCγ2 (Cell Signaling Technology), phosphorylated SRC Y416 (Cell Signaling Technology), total SRC (Cell Signaling Technology), phosphorylated paxillin Y118 (Cell Signaling Technology), total paxillin (Cell Signaling Technology), PARP, actin (Cell Signaling Technology).

    Article Title: Macrocyclic compounds and uses thereof
    Article Snippet: Protein lysates (approximately 20 μg) was resolved on 4-12% Bolt Bis-Tris precasted gels with MES running buffer (Life Technologies), transferred to nitrocellulose membranes using Trans-Blot Turbo Transfer System (Bio-Rad) and detected with antibodies targeting phosphorylated MET (Y1234/Y1235) (Cell Signaling Technology), MET (Y1349), MET (Y1003), total MET (Cell Signaling Technology), phosphorylated EGFR (Y1068) and total EGFR(Cell Signaling Technology), phosphorylated STAT3 and STAT5, total STAT3 and STAT5 (Cell Signaling Technology), phosphorylated AKT (Cell Signaling Technology), total AKT (Cell Signaling Technology), phosphorylated ERK (Cell Signaling Technology), total ERK (Cell Signaling Technology), phosphorylated PLCγ2 and total PLCγ2 (Cell Signaling Technology), phosphorylated SRC Y416 (Cell Signaling Technology), total SRC (Cell Signaling Technology), phosphorylated paxillin Y118 (Cell Signaling Technology), total paxillin (Cell Signaling Technology), PARP, actin (Cell Signaling Technology).

    Article Title: Chiral diaryl macrocycles and uses thereof
    Article Snippet: Protein lysates (approximately 20 μg) was resolved on 4-12% Bolt Bis-Tris precasted gels with MES running buffer (Life Technologies), transferred to nitrocellulose membranes using Trans-Blot Turbo Transfer System (Bio-Rad) and detected with antibodies targeting phosphorylated phosphorylated ROS1 and total ROS1 (Cell Signaling Technology), phosphorylated TRK A/B (Cell Signaling Technology), total TRKA antibody (Santa Cruz Biotechnology), phosphorylated SRC Y416 (Cell Signaling Technology), total SRC (Cell Signaling Technology), phosphorylated FAK Y576/577 (Cell Signaling Technology), total FAK (Cell Signaling Technology), phosphorylated paxillin Y118 (Cell Signaling Technology), total paxillin (Cell Signaling Technology), phosphorylated EGFR and total EGFR, CD44, and Tubulin (Sigma).



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    R&D Systems phosphorylated src
    FIGURE 2 Dose-dependent effects of chronic administration of clozapine on expression of pannexin1 and <t>phosphorylated</t> Src in the mPFC. Rats were chronically administered with clozapine (0, 5 and 10 mg kg1 day1 for 28 days). In the upper histograms, ordinate: mean ± SD (n = 6) of the relative levels of pannexin1 (PANX) per GAPDH (a) and phosphorylated-Src (pSrc) per Src (b). *P < 0.05: relative to control (clozapine free) by one-way ANOVA with Scheffe's post hoc test. F-values of the chronic clozapine administration on the expression of pannexin1 and pSrc in the rat frontal cortex using one-way ANOVA were [F(3,15) = 17.3 (P < 0.01)] and [F(3,15) = 18.0 (P < 0.01)], respectively. The lower panels indicate pseudo-gel images of capillary immunoblotting.
    Phosphorylated Src, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phosphorylated src y416 antibody
    FIGURE 2 Dose-dependent effects of chronic administration of clozapine on expression of pannexin1 and <t>phosphorylated</t> Src in the mPFC. Rats were chronically administered with clozapine (0, 5 and 10 mg kg1 day1 for 28 days). In the upper histograms, ordinate: mean ± SD (n = 6) of the relative levels of pannexin1 (PANX) per GAPDH (a) and phosphorylated-Src (pSrc) per Src (b). *P < 0.05: relative to control (clozapine free) by one-way ANOVA with Scheffe's post hoc test. F-values of the chronic clozapine administration on the expression of pannexin1 and pSrc in the rat frontal cortex using one-way ANOVA were [F(3,15) = 17.3 (P < 0.01)] and [F(3,15) = 18.0 (P < 0.01)], respectively. The lower panels indicate pseudo-gel images of capillary immunoblotting.
    Phosphorylated Src Y416 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/phosphorylated+src+y416+antibody/anti+phospho+src/us11981684-944-96-99
    Average 90 stars, based on 1 article reviews
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    Cell Signaling Technology Inc phosphorylated src y416
    rhMG53 inhibits FAK‐Src interaction and Src, Akt and ERK1/2 activation. A, HUVECs were stimulated with rhMG53 (10 μg/mL) or vehicle control for 24 h. Cell lysates were prepared and immunoprecipitated with anti‐FAK antibody. Then, captured proteins were analysed by Western blotting with anti‐Src and anti‐FAK antibodies as indicated. Representative images of 3 independent experiments are shown. B and C, HUVECs were exposed to rhMG53 (0, 5, 10 and 20 μg/mL) for 24 h. Phosphorylation of Src <t>Y416</t> , Akt T308 and ERK1/2, and total Src, Akt and ERK1/2 were analysed by Western blotting and representative images of 3 independent experiments are shown. The densitometric analysis of <t>phosphorylated</t> Src Y416 , Akt and ERE1/2 normalized to total Src, Akt and ERK1/2 was performed. All data shown are mean ± SD for 3 experiments and are expressed as fold changes. * P < .05; ** P < .01; *** P < .001
    Phosphorylated Src Y416, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore antibody against phosphorylated src at y416
    rhMG53 inhibits FAK‐Src interaction and Src, Akt and ERK1/2 activation. A, HUVECs were stimulated with rhMG53 (10 μg/mL) or vehicle control for 24 h. Cell lysates were prepared and immunoprecipitated with anti‐FAK antibody. Then, captured proteins were analysed by Western blotting with anti‐Src and anti‐FAK antibodies as indicated. Representative images of 3 independent experiments are shown. B and C, HUVECs were exposed to rhMG53 (0, 5, 10 and 20 μg/mL) for 24 h. Phosphorylation of Src <t>Y416</t> , Akt T308 and ERK1/2, and total Src, Akt and ERK1/2 were analysed by Western blotting and representative images of 3 independent experiments are shown. The densitometric analysis of <t>phosphorylated</t> Src Y416 , Akt and ERE1/2 normalized to total Src, Akt and ERK1/2 was performed. All data shown are mean ± SD for 3 experiments and are expressed as fold changes. * P < .05; ** P < .01; *** P < .001
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    R&D Systems phosphorylated src y416
    CLCa depletion significantly inhibits spreading-induced signaling. (A) siRNA-transfected cells held in suspension for 1 h (left panel) or plated on collagen IV-coated dishes for the indicated times (minutes, right panel) were lysed and subjected to western blotting with anti-active FAK [pFAK(Y397)], active Src <t>[pSrc(Y416)]</t> and anti-phosphorylated paxillin [pPax(Y118)] antibodies. (B) Protein phosphorylation in control cells at 30 min after plating was set as 100%. The results represent a summary from five to seven experiments. *P<0.05; **P<0.01. (C) Lysates from plated cells treated as in A were analyzed by western blotting with antibodies against Src-dependent FAK phosphorylation sites (Y576 and Y925). The blots shown represent one of three independent experiments.
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    FIGURE 2 Dose-dependent effects of chronic administration of clozapine on expression of pannexin1 and phosphorylated Src in the mPFC. Rats were chronically administered with clozapine (0, 5 and 10 mg kg1 day1 for 28 days). In the upper histograms, ordinate: mean ± SD (n = 6) of the relative levels of pannexin1 (PANX) per GAPDH (a) and phosphorylated-Src (pSrc) per Src (b). *P < 0.05: relative to control (clozapine free) by one-way ANOVA with Scheffe's post hoc test. F-values of the chronic clozapine administration on the expression of pannexin1 and pSrc in the rat frontal cortex using one-way ANOVA were [F(3,15) = 17.3 (P < 0.01)] and [F(3,15) = 18.0 (P < 0.01)], respectively. The lower panels indicate pseudo-gel images of capillary immunoblotting.

    Journal: British journal of pharmacology

    Article Title: Exploring the pathophysiology underlying clozapine-induced enhancement of glutamatergic transmission through L-glutamate and D-serine release associated with pannexin1 hemichannels.

    doi: 10.1111/bph.70112

    Figure Lengend Snippet: FIGURE 2 Dose-dependent effects of chronic administration of clozapine on expression of pannexin1 and phosphorylated Src in the mPFC. Rats were chronically administered with clozapine (0, 5 and 10 mg kg1 day1 for 28 days). In the upper histograms, ordinate: mean ± SD (n = 6) of the relative levels of pannexin1 (PANX) per GAPDH (a) and phosphorylated-Src (pSrc) per Src (b). *P < 0.05: relative to control (clozapine free) by one-way ANOVA with Scheffe's post hoc test. F-values of the chronic clozapine administration on the expression of pannexin1 and pSrc in the rat frontal cortex using one-way ANOVA were [F(3,15) = 17.3 (P < 0.01)] and [F(3,15) = 18.0 (P < 0.01)], respectively. The lower panels indicate pseudo-gel images of capillary immunoblotting.

    Article Snippet: Primary antibodies against GAPDH (NB300-327, RRID:AB_10001915, 1:300; Novus Biologicals, Littleton, CO, USA), pannexin1 (12595-1-AP, RRID:AB_ 2159186, 1:100; Proteintech, Rosemont, IL, USA), Src (2109, RRID: AB_2106059,1:50; Cell Signalling Technology, Danvers, MA, USA) and phosphorylated Src (pSrc; MAB2685, RRID:AB_3657975, 1 μg ml 1, R&D Systems, Minneapolis, MN, USA) were diluted in an antibody diluent (Immuno Shot Platinum; CosmoBio, Tokyo, Japan).

    Techniques: Expressing, Control, Western Blot

    FIGURE 6 Concentration-dependent effects of chronic administration of clozapine on expression of pannexin1 and phosphorylated Src in cultured astrocytes. (a and b) Concentration-dependent effects of chronic exposure to clozapine (3 and 10 μM for 28 days) on expression of pannexin1 and phosphorylated Src in the astroglial plasma membrane fraction, respectively. In the upper histograms, ordinate: mean ± SD (n = 6) of the relative levels of pannexin1 (PANX) per GAPDH or phosphorylated Src (pSrc) per Src. *P < 0.05: relative to control (clozapine free) by one- way ANOVA with Scheffe's post hoc test. F-values of the concentration-dependent effects of chronic clozapine administration on the expression of pannexin1 and pSrc using one-way ANOVA were [F(2,15) = 21.3 (P < 0.01)] and [F(2,15) = 14.9 (P < 0.01)], respectively. The lower panels indicate pseudo-gel images of capillary immunoblotting.

    Journal: British journal of pharmacology

    Article Title: Exploring the pathophysiology underlying clozapine-induced enhancement of glutamatergic transmission through L-glutamate and D-serine release associated with pannexin1 hemichannels.

    doi: 10.1111/bph.70112

    Figure Lengend Snippet: FIGURE 6 Concentration-dependent effects of chronic administration of clozapine on expression of pannexin1 and phosphorylated Src in cultured astrocytes. (a and b) Concentration-dependent effects of chronic exposure to clozapine (3 and 10 μM for 28 days) on expression of pannexin1 and phosphorylated Src in the astroglial plasma membrane fraction, respectively. In the upper histograms, ordinate: mean ± SD (n = 6) of the relative levels of pannexin1 (PANX) per GAPDH or phosphorylated Src (pSrc) per Src. *P < 0.05: relative to control (clozapine free) by one- way ANOVA with Scheffe's post hoc test. F-values of the concentration-dependent effects of chronic clozapine administration on the expression of pannexin1 and pSrc using one-way ANOVA were [F(2,15) = 21.3 (P < 0.01)] and [F(2,15) = 14.9 (P < 0.01)], respectively. The lower panels indicate pseudo-gel images of capillary immunoblotting.

    Article Snippet: Primary antibodies against GAPDH (NB300-327, RRID:AB_10001915, 1:300; Novus Biologicals, Littleton, CO, USA), pannexin1 (12595-1-AP, RRID:AB_ 2159186, 1:100; Proteintech, Rosemont, IL, USA), Src (2109, RRID: AB_2106059,1:50; Cell Signalling Technology, Danvers, MA, USA) and phosphorylated Src (pSrc; MAB2685, RRID:AB_3657975, 1 μg ml 1, R&D Systems, Minneapolis, MN, USA) were diluted in an antibody diluent (Immuno Shot Platinum; CosmoBio, Tokyo, Japan).

    Techniques: Concentration Assay, Expressing, Cell Culture, Clinical Proteomics, Membrane, Control, Western Blot

    FIGURE 7 Interaction among clozapine and inhibitors of Src and III-mGluR on the expression of pannexin1 and phosphorylated Src in the cultured astrocytes. (a) Interaction between 10 μM PP2 (Src inhibitor) and 10 μM clozapine on pannexin1 expression in the astroglial plasma membrane. (b) Interaction between 100 μM CPPG (III-mGluR inhibitor) and 10 μM clozapine on phosphorylated-Src expression in the astroglial plasma membrane. In the upper histograms, ordinate: mean ± SD (n = 6) of the relative levels of pannexin1 (PANX) per GAPDH or phosphorylated Src (pSrc) per Src. *P < 0.05: relative to control (clozapine free) by two-way ANOVA with Scheffe's post hoc test. @: P < 0.05: relative to 10 μM clozapine exposure by two-way ANOVA with Scheffe's post hoc test. F-values of the interaction between 10 μM PP2 and chronic exposure to 10 μM clozapine on the expression of pannexin1 using two-way ANOVA were [Fclozapine(1,20) = 55.5 (P < 0.01), FPP2(1,20) = 0.2 (P > 0.05), Fclozapine*PP2(1,20) = 1.1 (P > 0.05)]. F- values of the interaction between 100 μM CPPG and chronic exposure to 10 μM clozapine on the expression of pSrc using two-way ANOVA were [Fclozapine(1,20) = 56.0 (P < 0.01), FCPPG(1,20) = 27.7 (P < 0.01), Fclozapine*CPPG(1,20) = 0.4 (P > 0.05)]. The lower panels indicate pseudo-gel images of capillary immunoblotting.

    Journal: British journal of pharmacology

    Article Title: Exploring the pathophysiology underlying clozapine-induced enhancement of glutamatergic transmission through L-glutamate and D-serine release associated with pannexin1 hemichannels.

    doi: 10.1111/bph.70112

    Figure Lengend Snippet: FIGURE 7 Interaction among clozapine and inhibitors of Src and III-mGluR on the expression of pannexin1 and phosphorylated Src in the cultured astrocytes. (a) Interaction between 10 μM PP2 (Src inhibitor) and 10 μM clozapine on pannexin1 expression in the astroglial plasma membrane. (b) Interaction between 100 μM CPPG (III-mGluR inhibitor) and 10 μM clozapine on phosphorylated-Src expression in the astroglial plasma membrane. In the upper histograms, ordinate: mean ± SD (n = 6) of the relative levels of pannexin1 (PANX) per GAPDH or phosphorylated Src (pSrc) per Src. *P < 0.05: relative to control (clozapine free) by two-way ANOVA with Scheffe's post hoc test. @: P < 0.05: relative to 10 μM clozapine exposure by two-way ANOVA with Scheffe's post hoc test. F-values of the interaction between 10 μM PP2 and chronic exposure to 10 μM clozapine on the expression of pannexin1 using two-way ANOVA were [Fclozapine(1,20) = 55.5 (P < 0.01), FPP2(1,20) = 0.2 (P > 0.05), Fclozapine*PP2(1,20) = 1.1 (P > 0.05)]. F- values of the interaction between 100 μM CPPG and chronic exposure to 10 μM clozapine on the expression of pSrc using two-way ANOVA were [Fclozapine(1,20) = 56.0 (P < 0.01), FCPPG(1,20) = 27.7 (P < 0.01), Fclozapine*CPPG(1,20) = 0.4 (P > 0.05)]. The lower panels indicate pseudo-gel images of capillary immunoblotting.

    Article Snippet: Primary antibodies against GAPDH (NB300-327, RRID:AB_10001915, 1:300; Novus Biologicals, Littleton, CO, USA), pannexin1 (12595-1-AP, RRID:AB_ 2159186, 1:100; Proteintech, Rosemont, IL, USA), Src (2109, RRID: AB_2106059,1:50; Cell Signalling Technology, Danvers, MA, USA) and phosphorylated Src (pSrc; MAB2685, RRID:AB_3657975, 1 μg ml 1, R&D Systems, Minneapolis, MN, USA) were diluted in an antibody diluent (Immuno Shot Platinum; CosmoBio, Tokyo, Japan).

    Techniques: Expressing, Cell Culture, Clinical Proteomics, Membrane, Control, Western Blot

    FIGURE 9 Effects of chronic administration of L-BAIBA on Src signalling in the cultured astrocytes. (a and b) Interaction among chronic exposure to 30 μM L-BAIBA for 28 days and inhibitors of III-mGluR (100 μM CPPG) and NMDAR (1 μM MK801), respectively. In the upper side histogram, ordinate: mean ± SD (n = 6) of the relative levels of phosphorylated-Src (pSrc) per Src. *P < 0.05: relative to control (L-BAIBA free) by two-way ANOVA with Scheffe's post hoc test. @: P < 0.05: relative to 30 μM L-BAIBA exposure alone. F-values of the interaction between 100 μM CPPG and chronic exposure to 30 μM L-BAIBA on the expression of pSrc using two-way ANOVA were [FL-BAIBA(1,20) = 27.6 (P < 0.01), FCPPG(1,20) = 59.3 (P < 0.01), FL-BAIBA*CPPG(1,20) = 2.7 (P > 0.05)]. F-values of the interaction between 1 μM MK801 and chronic exposure to 30 μM L-BAIBA on the expression of pSrc using two-way ANOVA were [FL-BAIBA(1,20) = 46.3 (P < 0.01), FMK801(1,20) = 1.0 (P > 0.05), FL-

    Journal: British journal of pharmacology

    Article Title: Exploring the pathophysiology underlying clozapine-induced enhancement of glutamatergic transmission through L-glutamate and D-serine release associated with pannexin1 hemichannels.

    doi: 10.1111/bph.70112

    Figure Lengend Snippet: FIGURE 9 Effects of chronic administration of L-BAIBA on Src signalling in the cultured astrocytes. (a and b) Interaction among chronic exposure to 30 μM L-BAIBA for 28 days and inhibitors of III-mGluR (100 μM CPPG) and NMDAR (1 μM MK801), respectively. In the upper side histogram, ordinate: mean ± SD (n = 6) of the relative levels of phosphorylated-Src (pSrc) per Src. *P < 0.05: relative to control (L-BAIBA free) by two-way ANOVA with Scheffe's post hoc test. @: P < 0.05: relative to 30 μM L-BAIBA exposure alone. F-values of the interaction between 100 μM CPPG and chronic exposure to 30 μM L-BAIBA on the expression of pSrc using two-way ANOVA were [FL-BAIBA(1,20) = 27.6 (P < 0.01), FCPPG(1,20) = 59.3 (P < 0.01), FL-BAIBA*CPPG(1,20) = 2.7 (P > 0.05)]. F-values of the interaction between 1 μM MK801 and chronic exposure to 30 μM L-BAIBA on the expression of pSrc using two-way ANOVA were [FL-BAIBA(1,20) = 46.3 (P < 0.01), FMK801(1,20) = 1.0 (P > 0.05), FL-

    Article Snippet: Primary antibodies against GAPDH (NB300-327, RRID:AB_10001915, 1:300; Novus Biologicals, Littleton, CO, USA), pannexin1 (12595-1-AP, RRID:AB_ 2159186, 1:100; Proteintech, Rosemont, IL, USA), Src (2109, RRID: AB_2106059,1:50; Cell Signalling Technology, Danvers, MA, USA) and phosphorylated Src (pSrc; MAB2685, RRID:AB_3657975, 1 μg ml 1, R&D Systems, Minneapolis, MN, USA) were diluted in an antibody diluent (Immuno Shot Platinum; CosmoBio, Tokyo, Japan).

    Techniques: Cell Culture, Control, Expressing

    rhMG53 inhibits FAK‐Src interaction and Src, Akt and ERK1/2 activation. A, HUVECs were stimulated with rhMG53 (10 μg/mL) or vehicle control for 24 h. Cell lysates were prepared and immunoprecipitated with anti‐FAK antibody. Then, captured proteins were analysed by Western blotting with anti‐Src and anti‐FAK antibodies as indicated. Representative images of 3 independent experiments are shown. B and C, HUVECs were exposed to rhMG53 (0, 5, 10 and 20 μg/mL) for 24 h. Phosphorylation of Src Y416 , Akt T308 and ERK1/2, and total Src, Akt and ERK1/2 were analysed by Western blotting and representative images of 3 independent experiments are shown. The densitometric analysis of phosphorylated Src Y416 , Akt and ERE1/2 normalized to total Src, Akt and ERK1/2 was performed. All data shown are mean ± SD for 3 experiments and are expressed as fold changes. * P < .05; ** P < .01; *** P < .001

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: MG53 inhibits angiogenesis through regulating focal adhesion kinase signalling

    doi: 10.1111/jcmm.16777

    Figure Lengend Snippet: rhMG53 inhibits FAK‐Src interaction and Src, Akt and ERK1/2 activation. A, HUVECs were stimulated with rhMG53 (10 μg/mL) or vehicle control for 24 h. Cell lysates were prepared and immunoprecipitated with anti‐FAK antibody. Then, captured proteins were analysed by Western blotting with anti‐Src and anti‐FAK antibodies as indicated. Representative images of 3 independent experiments are shown. B and C, HUVECs were exposed to rhMG53 (0, 5, 10 and 20 μg/mL) for 24 h. Phosphorylation of Src Y416 , Akt T308 and ERK1/2, and total Src, Akt and ERK1/2 were analysed by Western blotting and representative images of 3 independent experiments are shown. The densitometric analysis of phosphorylated Src Y416 , Akt and ERE1/2 normalized to total Src, Akt and ERK1/2 was performed. All data shown are mean ± SD for 3 experiments and are expressed as fold changes. * P < .05; ** P < .01; *** P < .001

    Article Snippet: Antibodies to phosphorylated FAK Y397 , FAK, phosphorylated Src Y416 , Src, phosphorylated Akt T308 , Akt, phosphorylated ERK1/2 and ERK1/2 were obtained from Cell Signaling Technology.

    Techniques: Activation Assay, Control, Immunoprecipitation, Western Blot, Phospho-proteomics

    CLCa depletion significantly inhibits spreading-induced signaling. (A) siRNA-transfected cells held in suspension for 1 h (left panel) or plated on collagen IV-coated dishes for the indicated times (minutes, right panel) were lysed and subjected to western blotting with anti-active FAK [pFAK(Y397)], active Src [pSrc(Y416)] and anti-phosphorylated paxillin [pPax(Y118)] antibodies. (B) Protein phosphorylation in control cells at 30 min after plating was set as 100%. The results represent a summary from five to seven experiments. *P<0.05; **P<0.01. (C) Lysates from plated cells treated as in A were analyzed by western blotting with antibodies against Src-dependent FAK phosphorylation sites (Y576 and Y925). The blots shown represent one of three independent experiments.

    Journal: Journal of Cell Science

    Article Title: A unique role for clathrin light chain A in cell spreading and migration

    doi: 10.1242/jcs.224030

    Figure Lengend Snippet: CLCa depletion significantly inhibits spreading-induced signaling. (A) siRNA-transfected cells held in suspension for 1 h (left panel) or plated on collagen IV-coated dishes for the indicated times (minutes, right panel) were lysed and subjected to western blotting with anti-active FAK [pFAK(Y397)], active Src [pSrc(Y416)] and anti-phosphorylated paxillin [pPax(Y118)] antibodies. (B) Protein phosphorylation in control cells at 30 min after plating was set as 100%. The results represent a summary from five to seven experiments. *P<0.05; **P<0.01. (C) Lysates from plated cells treated as in A were analyzed by western blotting with antibodies against Src-dependent FAK phosphorylation sites (Y576 and Y925). The blots shown represent one of three independent experiments.

    Article Snippet: Antibodies against the following proteins were used: CLCa (1:1000, sc-28276), CLCb (1:500, sc-376414), actin (1:1000, sc-1616) from Santa Cruz Biotechnology, FAK (1:2000, 610088) and β1-integrin (1:1000, 610467) from BD Transduction Labs, phosphorylated FAK(Y397) (1:1000, 44-624G), phosphorylated paxillin(Y118) (1:1000, 44-722G) from Fisher Scientific, Src (1:2000, 2108), phosphorylated Src(Y416) (1:1000, MAB2685, 2101), phosphorylated FAK(Y576) (1:1000, 3281), FAK(Y925) (1:1000, 3284) from Cell Signaling, phosphorylated Src(Y416) (1:1000, MAB2685) from RD Systems, WAVE1/Scar (1:1000, 07-037), Rac1 (1:2000, 05-389) from Millipore.

    Techniques: Transfection, Western Blot