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Figure 2


Fig. 2. Ang II-mediated phosphorylation of β2-adaptin involves Src. (A) COS-7 cells transfected with HA-AT1R and Flag-β2-adaptin were incubated for 30 minutes with either the vehicle (DMSO), the Src inhibitor (PP2, 4 µM), the EGFR inhibitor (PD158780, 50 nM) or the less active form of PP2 (PP3, 4 µM), before adding Ang II (1 µM). Flag-β2-adaptin was immunoprecipitated using an anti-Flag antibody, and the immunoprecipitates were analyzed by western blot as described in Materials and Methods. Data are presented as the mean ± s.e.m. of three independent experiments. They represent the percent of β2-adaptin phosphorylation as compared to vehicle, after normalizing for equal amounts of β-adaptin. ** indicates P<0.01 vs DMSO determined by one-way ANOVA, and was considered significant. (B) COS-7 cells transfected with AT1R, Flag-β2-adaptin and either Src wild type or Src K298R (HA-Src and HA-Src K298R) were left untreated (–) or stimulated (+) with Ang II (1 µM). Detection of the phosphorylated β2-adaptin in the immunoprecipitates was performed as described in (A).





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