Fig. 8

48 kDa PTPN2 forms a signaling complex with PKD and Akt. HEK-PAFR were transiently transfected with D182A-48 kDa or − 45 kDa PTPN2 constructs tagged with Flag or control vector (Ctrl). Cells were incubated overnight in DMEM-0.2% BSA, then stimulated with PAF (100 nM) or vehicle for indicated times. Stimulation was stopped on ice, cells were scraped and lysed. Whole cell lysates were submitted to immunoprecipitation using anti-FLAG antibodies coupled to agarose beads. a Precipitates were separated by SDS-PAGE, transferred to nitrocellulose membranes and blotted overnight with anti-Akt, anti-PKD or anti-Flag antibodies. Lysate samples have been loaded onto the same blots as immunoprecipitated PTPN2 samples and serve as loading controls. a Representative blots and b compilations of at least 3 independent experiments are shown. The data presented are mean ± SEM. Significance was established with two-way ANOVA with Sidak post-test. *p < 0.05