Figure 1. H₂O₂-induced activation of Src and c-Abl and resultant phosphorylation of Cav-1.

(A) H₂O₂ induced the activation of Src and phosphorylation of Cav-1 in a concentration-dependent manner. RLMVECs were exposed to different concentrations of H₂O₂ (0.01-0.8 mmol/L) for 30 min. (B) Time course of H₂O₂-induced Cav-1 phosphorylation. Cells were stimulated with 0.6 mmol/L H₂O₂ for indicated times. (C) Effect of PP2 on Cav-1 phosphorylation induced by different concentrations of H₂O₂. (D) Dose-dependent c-Abl phosphorylation induced by H₂O₂. Cells were exposed to vehicle or H₂O₂ (0.1-0.6 mmol/L) for 30 min. c-Abl phosphorylation was measured by immunoprecipitation with c-Abl antibody and immunoblotting with phosphotyrosine antibody. Immunoblot of total c-Abl is shown as a loading control. (E) Effect of c-Abl siRNA on Cav-1 phosphorylation induced by the different concentrations of H₂O₂. (F) Effect of PP2 on H₂O₂-stimulated c-Abl activation. Cells were pretreated with PP2 or vehicle for 15 min and then stimulated with H₂O₂ (0.6 mmol/L) for 30 min. All blots are representative of 3 separate experiments.