Fig. 7.
TSP1 increases ZA protein tyrosine phosphorylation through EGFR/ErbB2 activation. HMVEC-Ls were incubated for 1 h with TSP1 (30 μg/ml, i.e., 214 nM) or medium alone, in the presence or absence of either AG1478 (5 μM) or AG825 (5 μM), as well as in the presence of vanadate (200 μM) and PAO (1.0 μM), only during the last 0.25 h of incubation. ECs were lysed and immunoprecipitated with antibodies raised against VE-cadherin (A), γ-catenin (B), or p120ctn (C). The immunoprecipitates were resolved by SDS-PAGE and transferred to PVDF, and the blots were probed with antiphosphotyrosine antibody. Blots were stripped and reprobed with the immunoprecipitating antibodies for normalization of phosphotyrosine signal to the loaded immunoprecipitated protein. These blots are representative of 2 independent experiments.