Fig. 4
PTP1B increases PAF-induced GSK-3 phosphorylation. a iMo-DCs collected on day 7 were incubated at 106 cells/ml in RPMI+ 0.2% BSA for 5 h, then incubated for 20 min with 10 μM PTP1B inhibitor before being stimulated with 10 nM PAF for indicated times. Reaction was stopped on ice and cells were collected and lysed. b siCTRL-MoDCs or siPTP1B-MoDCs were collected on day 7, incubated at 106 cells/ml in RPMI+ 0.2% BSA for 5 h and stimulated for indicated times with 10 nM PAF before being lysed. a and b Lysates were separated on SDS-PAGE gels and Western blots were performed with anti-vinculin, anti-pSer21/Ser9 GSK-3 and anti-GSK-3ß Abs. The blots were stripped between re-blotting with indicated antibodies. Representative blots are shown for each experimental condition and compilations of at least 3 independent experiments are presented as mean ± S.E.M of normalized ratios of pSer21/9 GSK-3 calculated as described in Methods. Significance was established with paired two-way ANOVA with Sidak post-test: *p < 0.05, **p < 0.01 vs control cells