Sorafenib affects intracellular signaling pathways. LPS (100 ng/mL) was added to immature DCs 15 minutes (A, top figure) or 24 hours (A, bottom figure, B-E) before harvesting. Protein levels were detected by SDS-PAGE and Western blot. Ponceau S staining was performed to ensure equal loading of the gel. (A) Sorafenib affects the TLR pathway. MyD88 and TRAF6 levels were reduced by sorafenib treatment. (B) Sorafenib reduces nuclear localization of the NF-κB family members c-Rel, Rel-B, Rel-A, and of PU.1 and IRF-3 in mature cells and nuclear translocation of Rel-B, IRF-3, and PU.1 also in immature cells. Protein levels of IRF-8 in nuclei were not affected. (C) The inhibitory effects of sorafenib partly are mediated by MAPK cascade and the PI3K/AKT pathway. Sorafenib reduces phosphorylation states of p38, whereas sorafenib induces phosphorylation of ERK. (D) Addition of sunitinib to the culture medium has no effect on the signaling pathways in DCs. (E) Addition of sunitinib to the culture medium has no effect on nuclear translocation of c-Rel, Rel-B, Rel-A, PU.1, IRF-3, and IRF-8. Representative experiments of at least 4 are presented.