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. 2022 Feb 22;71(5):1149–1165. doi: 10.2337/db21-0830

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© 2022 by the American Diabetes Association

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VEGF-mediated increase in endothelial cell functionality is PLCγ2 dependent. OCR (A) and ECAR (B) calculation of HMECs in normal conditions and after VEGF exposure (100 ng/mL) and cotreatment with either control (con) siRNA or PLCγ2 siRNA (SI) using the Seahorse XF 96-well plate reader. OCR was determined during sequential treatments with oligomycin (ATP synthase inhibitor), carbonyl cyanide-4 (trifluoromethoxy) phenylhydrazone (FCCP), a protonophore that lowers the mitochondrial membrane potential to create conditions for maximal oxidative respiration, and antimycin A plus rotenone to inhibit the electron transport chain. C: Basal respiration, maximal respiration, proton leak, and nonmitochondrial (non-mito) respiration measurement in HMECs in normal conditions and supplemented with VEGF (100 ng/mL) and cotreatment with control (con) siRNA or PLCγ2 siRNA (SI). Matrigel tube formation images (D) and tube length (E) analysis in HMECs in normal conditions and after cotreatment with control (con) siRNA or PLCγ2 siRNA. Scale bars, 200 µm. n = 6. *P < 0.05 (one-way ANOVA, followed by Tukey HSD post hoc test).