Fig. 3. β-arrestin–AP-2–Src complex is involved in β2-adaptin phosphorylation. (A,B) HEK293 cells were transfected with AT1R, Flag-β2-adaptin and either pcDNA3.1, HA-SH1 or HA-SH1-KD (A) or with AT1R and either Flag-β2-adaptin wild type or Flag-β2-adaptin E902A (B), before either being left untreated (–) or stimulated (+) with Ang II (1 µM). Flag-β2-adaptin were immunoprecipitated from cell lysates using an anti-Flag antibody, and the immunoprecipitates were analyzed by western blot as previously described. Whole cell extracts (Total: HA) were probed for the detection of HA-SH1 and HA-SH1-KD using an anti-HA antibody (A). Data are presented as the mean ± s.e.m. for three independent experiments.