Specific inhibition of band-3 coimmunoprecipitation in SHP-2 IPs by P-Y359–containing phosphopeptide.

Erythrocytes were stimulated with 2 mM diamide. Extracts from cell membranes were immunoprecipitated with anti–SHP-2 antibody in the absence (lane 1) or presence (lanes 2-5) of increasing concentrations of synthetic phosphopeptides reproducing band-3 sequences containing either P-Y359 (A) or P-Y904 (B). SHP-2 IPs were submitted to SDS-PAGE, transferred to nitrocellulose, and immunostained with anti–band 3 antibody. Blots were then stripped and reprobed with anti–SHP-2 antibody. Figure is representative of at least 3 separate experiments.