Fig. 8

PAF increases interaction between PTP1B and Src. HEK-PAFR were transiently transfected with D181A PTP1B tagged with Flag or control vector expressing Flag alone (CV). Cells were incubated overnight in DMEM-0.2% BSA, then stimulated with PAF for indicated times. Stimulation was stopped on ice, cells were scraped and lysed. Whole cell lysates were immunoprecipitated using anti-Flag Abs coupled to agarose beads. Precipitates were loaded onto SDS-PAGE, transferred to nitrocellulose membranes and blotted overnight with anti-Src and then with anti-Flag Abs. The doted lines indicate where an empty well was removed in order to align the wells with the appropriate wells in the lysate. Representative blots and compilations of 3 independent experiments are shown. The data presented are mean ± S.E.M of normalized Src values (over PTP1B) over values for unstimulated cells