TLR4 signaling and the inhibition of liver hepcidin expression by alcohol

Figure 3 Nuclear factor-κB phosphorylation. Cytosolic (A, C) and nuclear (B, D) protein fractions isolated from the livers of C3H/HeJ and C3H/HeOuJ mice pair-fed with regular and ethanol-containing diets were used for western blots to detect phosphorylated (serine 536) p65 (p-p65) expression. Anti-total p65 (p65) (C) and anti-TATA-binding protein (TBP) (D) antibodies were used as controls. Cytosolic (E) and nuclear (F) p-p65 expression was quantified by densitometric analysis and normalized to total p65 or TBP protein expression, respectively. Normalized p-p65 expression in ethanol-fed mice was expressed as fold expression of that in the respective mice pair-fed with the regular diet (mean; n = 3 per group).