Treatment of human erythrocytes induces both Tyr phosphorylation of band 3 and translocation of cytosolic SHP-2 to membranes.

Red blood cells were incubated in the absence (lane 1) or presence of indicated amounts of pervanadate (lanes 2-4), diamide (lanes 5-7), and NEM (lanes 8-10). Cell membranes were isolated, solubilized, and submitted to SDS-PAGE followed by transfer to nitrocellulose, as described in “Materials and methods.” The upper part of blots containing band 3 (101.8 kd) was immunostained with anti–P-Y antibody (B), stripped, and reprobed with anti–band 3 antibody (A). The lower part of blots containing SHP-2 (68 kd) was immunostained with anti–SHP-2 antibody (C). Panels are representative of at least 7 separate experiments.