Fig. 3
PTP1B decreased PAF-induced C/EBPß phosphorylation levels. a-c iMo-DCs, collected on day 7, were incubated at 106 cells/ml in RPMI+ 0.2% BSA for 5 h. Cells were then incubated for 20 min with 10 μM PTP1B inhibitor before being stimulated with 10 nM PAF for indicated times. d & e siCTRL-MoDCs or siPTP1B-MoDCs were collected on day 7 and incubated at 106 cells/ml in RPMI+ 0.2% BSA for 5 h. Cells were then stimulated for indicated times with 10 nM PAF. a-e Reaction was stopped on ice and cells were collected and lysed. Lysates were separated on SDS-PAGE gels and Western blots were performed with anti-vinculin, anti-pThr235 C/EBPß (LAP1 and LAP2), which also recognizes pThr37 LIP, and anti-C/EBPß antibodies (Abs). a A replicate blot is shown with anti-C/EBPβ antibodies (Abs) and anti-vinculin. d The blots were stripped between re-blotting with indicated antibodies. Representative blots are shown for each condition and compilations of at least 3 independent experiments are presented as mean ± S.E.M of normalized ratios pTh235 C/EBPß levels calculated as described in Methods. Significance was established with paired two-way ANOVA with Sidak post-test: *p > 0.05, **p > 0,01 over control cells