Effects of ATII on phosphorylation of EGFR.

<p>A, HEK293 cells stably transfected with AT_1AR were serum starved for 5 hours followed by 100nM ATII for 10 min. Phosphorylation of EGFR on Thr-654 (P-Thr654) was determined by western blotting. Blots were stripped and reprobed for total EGFR to normalize for loading. B, HEK293 cells stably transfected with AT_1AR were serum starved for 5 hours and treated with 5ng/ml of EGF for 5 min with or without 1 hour pretreated with 100nM ATII. Phosphorylation of EGFR on Tyrosine 1045 (P-Tyr1045) was determined by western blotting. The blots were stripped and reprobed for total EGFR to normalize for loading. C, HEK293 cells stably transfected with AT_1AR were serum starved for 5hours and then pretreated with Gö6976, FIPI or vehicle followed with 100nM ATII or vehicle for 1 hour and then treated with 5ng/ml EGF for 5 min. P-Tyr1045 and EGFR were determined by western blotting. D, HEK293 cells stably transfected with AT_1AR were serum starved for 5hours and then pretreated with Gö6976, 1-butanol or vehicle followed with 100nM ATII or vehicle for 5min, P-Tyr1068 and EGFR were determined by western blotting. For all figures, * p <0.05, ***p <0.001 from at least three independent experiments.