FIG. 8.

Mutation of dynamin's Src-phosphorylated tyrosines reduces dynamin's ability to downregulate Pyk2 Y402 phosphorylation. 293VnR cells were cotransfected with Pyk2 and either wild-type dyn-GFP, the Src phosphorylation site mutants dynY231F-GFP and dynY597F-GFP, or empty vector as indicated. Pyk2 was immunoprecipitated, and the immune complexes were analyzed for Pyk2 by Western blotting. Membranes were stripped and reblotted for phospho-Y402. Pyk2 was also immunoprecipitated and blotted with dynamin and Pyk2. TCLs were blotted for transfected dyn-GFP. Experiments were reproduced four times, and the results of a representative experiment are shown. Labels: IP, immunoprecipitation; B, Western blot; Dyn, dynamin.