Figure 3.

Pulse–chase analysis of wild-type and mutant Src proteins. Src−/− MEFs were transfected with pLNCX vector expressing c-Src, or the following Src mutants: Y527F, E378G, Y416F, or K295R. Forty eight hours posttransfection, cells were pulse-labeled with 100 μCi ³⁵S Met/Cys for 45 min. Plates were incubated in “chase” media with 100-fold excess methionine and then harvested at the indicated times. Lysates were immunoprecipitated by using mAb 327 and separated by SDS/PAGE. Gels were quantitated with a PhosphorImager, and each time point was normalized to the 0-hr time point.