Levels of p65 and p50 NF-κB subunits and IKKβ protein were examined by western blotting. Each protein analyzed was identified as a single band with the predicted molecular size–65 (p65), 50 (p50) and 87 (IKKβ) kDa (Figure 1). The optical density values obtained for the proteins were within the linear range of detection as was evident from experiment with serial dilutions of reference samples; densitometry assessment of bands showed linear dependence with correlation coefficient equal to 0.998 between protein immunoreactivity measured as optical density and protein load measured as Memcode optical density (see also [37]). For inter-gel comparison, the optical density of proteins in each sample was expressed as a percentage of reference samples on a given gel, and these values were used for statistical analysis. Similar to the levels of RELA mRNA, the p65 levels in the PFC of alcoholics were decreased by 1.46-fold (Table 2) but the differences did not reach statistical significance. The levels of p65 in the MC, and those of p50 and IKKβ proteins in neither the PFC (Table 2; n = 15) nor MC
