Fig 4.
SHP-2 dimerization is mediated by a disulfide bridge. T-Rex SHP-2 293 cells were transfected with SHP-2_V5 phosphatase, and expression of SHP-2_FLAG was induced for 6 h with doxycycline. The SHP-2_FLAG protein and its molecular partners were immunoprecipitated with anti-FLAG antibody and then treated with DTT. The asterisk indicates a band with an Mr of about 130,000 that was visible only when DTT was omitted in the loading dye. Immunoblots are from one experiment representative of three that gave similar results. (B) T-Rex SHP-2 293 cells were transfected with SHP-2_V5 phosphatase induced with doxycycline. The lysate was split in two, and one half was treated with DTT for 15 min (lane 2). The SHP-2_FLAG protein was immunoprecipitated, and the interaction was revealed by probing with anti-V5 antibody.