Figure 3

C-terminal end of RhoGDIα is tyrosine phosphorylated. (A) RhoGDIα was expressed alone or with pEGFP-tagged Fer as indicated above the panel. Cells were stimulated with pervanadate (0.5 mM H₂O₂ + 0.5 mM vanadate) (+) and RhoGDIα was immunoprecipitated. pY20 blots were stripped and re-probed with GFP or RhoGDIα antibodies. Antibodies used for detection are indicated below the panels. (B) Xpress-tagged C-terminal RhoGDIα (CT) was expressed alone or together with pEGFP-tagged Fer. Cells were stimulated with pervanadate for 5 minutes. Results are representative of three independent experiments. (C). RBL-2H3 cells were treated with pervanadate for 5 minutes (+) as indicated and endogenous RhoGDIα was immunoprecipitated. Western blots were reacted with antibodies against RhoGDIα or phosphotyrosine. The RhoGDIα antibody was from Santa Cruz.