FIGURE 5.

VEGF-induced ACC phosphorylation but not eNOS phosphorylation is inhibited in endothelial cells from AMPKα1^−/− mice. MLEC from AMPKα1^+/+ and AMPKα1^−/− mice were stimulated with 50 ng/ml VEGF for the indicated times. Cell lysates were subjected to Western blot analysis using antibodies against phosphorylated AMPKα (Thr¹⁷²), AMPKα1, or pan-AMPKα (A); phosphorylated (serine 1177) or total eNOS (B); and phosphorylated or total ACC (C). Representative blots and densitometric analysis from two experiments for each treatment are shown (mean ± S.E.). In addition to the kinetic studies shown here, three experiments with a single VEGF stimulation (5 min) were performed. Statistical analysis for the 5-min time points is given under “Results”. A significant inhibition of ACC but not of eNOS phosphorylation was observed.