Fig. 9

48 kDa PTPN2 modulates Akt activation via a PKD-dependent pathway. a, b HEK-PAFR were transiently transfected with 48 kDa or 45 kDa PTPN2 constructs tagged with FLAG, PKD tagged with GST or control vectors (pcDNA3 or pcDNA3-GST). 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 separated by SDS-PAGE, transferred to nitrocellulose membranes and blotted overnight with anti-pSer473Akt, anti-GST, anti-actin, anti-FLAG and anti-Akt antibodies. Results shown are a compilation of 3 independent experiments, expressed as variations of Akt phosphorylation due to PTPN2 overexpression where values of pAkt levels of control cells (without PTPN2) are set to 1 (Dotted line) for each indicated stimulation time. c HEK-PAFR were transiently co-transfected with the hIL-6-luc, control vector pcDNA3, 48 kDa PTPN2 or PKD constructs. Cells were incubated overnight in DMEM-0.2% BSA and stimulated with PAF (100 nM) or vehicle for 6 h and luciferase activity was measured. The data presented are mean ± SEM of at least 3 experiments performed in triplicate. a–c The data presented are mean ± SEM. Significance was established with two-way ANOVA with Sidak post-test. *p < 0.05