Figure 4.

Effect of SHP-2(C/S) on phosphorylation and association of proteins with Gads and SLP-76. Jurkat T cells were transiently transfected with an empty vector (Vector) or an expression vector encoding SHP-2(C/S) or SHP-2 WT. Cells were stimulated for 5 min by anti-CD3 crosslinking. Lysates were immunoprecipitated with (A) anti-Gads, (B) anti-SLP-76, (C) anti-Vav1 or (D) anti-ADAP antibody. Immunoprecipitates were separated by SDS–PAGE, immunoblotted for phosphotyrosine and then stripped and reprobed for the indicated proteins. (C) Anti-Vav1 immunoprecipitates were also blotted with a phosphospecific antibody to Vav1 phosphorylated at Y174.