Figure 4.

Both P424 and P467 of Akt are required for the association of PLC-γ1 and Akt. (A) COS-7 cells were transfected with GFP-Akt or GFP-tagged Akt mutants P424A, P467A, and P/P. The cells were either not treated or treated with 100 ng/ml EGF for 30 min. After treatment, the cell lysates were immunoprecipitated with anti-PLC-γ1 antibody, and the resulting immunoprecipitates were immunoblotted with a polyclonal anti-GFP antibody. The PLC-γ1 blot served as the loading control, and the GFP blot of total lysate served as the control of expression. (B) COS-7 cells were transfected with GFP-Akt-P/P. The cells were then fixed and stained with anti-Akt antibody. The localization of Akt was indicated by arrow. Bar, 20 μM.