Fig. 6

VEGFR2 association with caveolin-1 is markedly decreased by treatment with VEGF, cyclodextrin, or filipin and is partially rescued by treatment with cholesterol repletion. VEGFR2 was immunopricipitated from PBECs lysates, and its association with caveolin-1 was assessed by Western blot. Actin in the supernatant was probed to ensure equal immunoprecipitation across conditions (A). PBECs were treated with VEGF (50 ng/ml) in the presence or absence of pretreatment with the caveolar-disrupting agent cyclodextrin (CD; 5 mM, 30 min) or filipin (2.5 g/ml, 30 min). Reversal of drug effects was sought with simultaneous cholesterol repletion (Chol; 15 g/ml) given at the time of cyclodextrin administration. Caveolin-1was immunopricipitated from PBECs lysates, and its association with VEGFR2 was assessed by Western blot. Actin in the supernatant was probed to ensure equal immunoprecipitation across conditions (B). All data are representative of three independent experiments