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. 2023 Oct 16;133(20):e161366. doi: 10.1172/JCI161366

Figure 6. TG rescues activation of Src and VE-cadherin phosphorylation in response to VEGFA, after removal of PLCγ.

Figure 6

(A–C) PLA using antibodies against Src and pSFK Y418, visualizing phosphorylation of Src on Y418, in HUVECs treated for 5 min with DMSO, 100 ng/ml VEGFA or 100 ng/ml VEGFA + 1uM TG, and pretreated with siCtr (A), siPLCG1 (B), or siNOS3 (C). Junctions are stained for VE-cadherin (VEC) and nuclei with DAPI (blue). Scale bar: 20 μm. Boxed regions in left panels are magnified in panels to the right. Scale bar: 5 μm. (D) MFI quantifications of junctional PLA signals representing Src phosphorylated on Y418 from A–C; n = 3 independent experiments, ≥ 3 fields of view/experiment. 1-way ANOVA. (E) Representative images of immunofluorescent stainings with antibodies against VEC and pVEC Y685, of HUVECs treated with VEGFA or VEGFA+TG, pretreated with siCtr, siPLCG1 or siNOS3. Scale bar: 20 μm. (F) MFI quantification of data from E shown as fold of DMSO-treated siCtr; n = 3 independent experiments, ≥ 3 fields of view/experiment. 1-way ANOVA. Data represent mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001.