FIG. 6.
In vivo association and tyrosine phosphorylation of PLC-γ by NPM-ALK require Tyr664. (A) Coprecipitation of PLC-γ with NPM-ALK was examined in lysates of 5 × 107 Ba/F3 cells stably transfected with wt NPM-ALK (WT) and NPM-ALK(Y664F). Immunoprecipitation was performed with anti-ALK antibody, rabbit anti-mouse antibody (Con.), or anti-PLC-γ antibody. Samples were resolved by SDS–7.5% PAGE and analyzed by immunoblotting with anti-ALK antibody. (B) Expression levels of PLC-γ (left panel) and tyrosine phosphorylation of NPM-ALK and PLC-γ (right panel) in Ba/F3 cells transfected with wt NPM-ALK and NPM-ALK(Y664F) were determined by immunoprecipitation and immunoblotting with the antibodies indicated. IP, immunoprecipitation; WB, Western blotting.