FIG. 8.

Analysis of multisite “pocket” mutants of c-Src. (A) COS-1 cells transfected with WT and mutant c-Src constructs were lysed at 22 or 48 h posttransfection, and lysates were analyzed by Western blotting with the indicated antibodies. (B) COS-1 cells transfected with WT or mutant c-Src constructs were labeled with 13-[¹²⁵I]iodotridecanoic acid, and radiolabel incorporation into c-Src was analyzed as described in the legend to Fig. 7C. (C) Lysates from COS-1 cells cotransfected with NMT and WT or mutant c-Src were subjected to cell fractionation and analyzed as described in the legend to Fig. 1C. (D) COS-1 cells transfected with WT or mutant c-Src were analyzed at 48 h posttransfection for c-Src expression, using indirect immunofluorescence. (E) COS-1 cells transfected with WT or mutant c-Src were lysed at 24 h posttransfection. c-Src was immunoprecipitated and used in an in vitro kinase assay with purified Cas. Combined data from two independent experiments are shown. Error bars indicate standard deviations. The inset shows that equal amounts of WT and mutant Src were immunoprecipitated.