Fig. 1.

Effect of VEGF on AMPK activity and eNOS Ser1177 phosphorylation. (A) HAECs were incubated with 10 ng/ml VEGF for various times and lysates prepared. (B) HAECs were incubated with the indicated concentrations of VEGF for 5 min and lysates prepared. Total AMPK was immunoprecipitated from endothelial cell lysates and assayed for AMPK activity. In parallel experiments, lysates were subjected to SDS–PAGE, transferred to nitrocellulose and probed with the antibodies indicated. The intensity of the resultant bands was quantified using NIH Image software. Representative immunoblots are shown, repeated with similar results on three different samples of lysates. The results are expressed as the means ± SD % basal AMPK activity (■) or % basal eNOS phosphorylation (○) for three independent experiments. *p < 0.01 relative to value in absence of VEGF. ^#p < 0.05 relative to value in absence of VEGF.