Figure 3: Smad2 Activation. The phosphorylation of Smad2 protein in the liver lysates of mice fed with regular (control) or ethanol-containing (alcohol) Lieber-De Carli liquid diets was determined by western blotting employing an anti-phospho-Smad2 antibody, as described in Materials and Methods. (b) An antibody recognizing the total levels of Smad2 protein was employed as a control to confirm equal protein loading. (c) Autoradiographs from different experiments ( ) were scanned by a densitometer, and phospho-Smad2 (p-Smad2) expression in each sample was quantified by normalizing to total Smad2 protein expression. Normalized phospho-Smad2 expression in alcohol-treated mice was expressed as fold expression of that in control mice.