Fig. 12

The Akt pathway modulates GSK-3 and C/EBPß but only at later stimulation times. iMo-DCs were collected on day 7 and incubated for 20 min with 2.5 μM Akt inhibitor MK-2206 or with its vehicle (DMSO). Cells were then stimulated with 10 nM PAF for indicated times. Reaction was stopped on ice, cells were collected and lysed. Lysates were separated on SDS-PAGE gels and Western blots were performed with Abs recognizing vinculin and a pSer21/9 GSK-3 and GSK-3ß or b pThr235 C/EBPß and C/EBPß. The blots were stripped between re-blotting with indicated antibodies. The doted lines indicate where the image was slightly rotated for better alignment with the rest of the blot, due to warped migration. Representative blots are shown for each experimental condition and compilations of experiments are presented as mean ± S.E.M of normalized ratios of phospho-protein levels for at least 4 experiments. Significance was established with paired two-way ANOVA with Sidak post-test: *p < 0.05 vs control cells