Figure 2. Functional inhibition of SYK inhibits alcohol-induced pSYKY525/526, NF-κB nuclear binding, and pro-inflammatory mediators.
C57BL/6 mice were fed a chronic alcohol or an isocaloric pair fed (PF) diet with SYK inhibitor (SYK Inh.) treatment or vehicle treatment (+ Veh.). Total liver protein was analyzed for activated pSYKY525/526 by western blot (A). NF-κB nuclear binding was assessed from liver nuclear extracts by electrophoretic mobility shift assay (B). Total liver RNA and protein was assessed by RT-qPCR and ELISA for TNF-α mRNA and protein, respectively (C). Total liver mononuclear cell protein was analyzed for activated pSYKY525/526 by western blot (D). Total liver protein and liver samples were analyzed for F4/80 by western blot (E), CD68 by immunohistochemistry (E) and iNOS protein by western blot (F). A p-value <0.05 was considered statistically significant by ANOVA for 7–10 mice per experimental group (*, p<0.05 compared to baseline; #, p<0.05 compared to the EtOH + vehicle group).