Fig. 4. Ang2 activates a panel of non-receptor protein tyrosine kinases. (A,B) IBE cells (6 cm dishes) were either stimulated or left unstimulated with 1 µg/ml Ang2 for 15 minutes and c-Fyn (A) or c-Src (B) was immunoprecipitated from 90% of lysate and used for in vitro kinase assay. Acid-denatured enolase was used as a substrate. The remaining 10% total lysate was examined by immunoblotting to determine the amount of loaded proteins. (C) IBE cells expressing FLAG-tagged wild-type c-Fes (FesWT6-8 cells; 10 cm dishes) were either stimulated or left unstimulated with 1 µg/ml Ang2 for 15 minutes and FLAG-tagged c-Fes was immunoprecipitated followed by imunoblotting. Between two probings, membranes were stripped. Representative data were obtained from two independent experiments.