Alcohol Activates TGF-Beta but Inhibits BMP Receptor-Mediated Smad Signaling and Smad4 Binding to Hepcidin Promoter in the Liver
Figure 8
Smad4 DNA-Binding Activity. Smad DNA-binding activity was determined by electrophoretic mobility gelshift assays (EMSA), as described in Materials and Methods. 7 μg of nuclear lysate protein isolated from the livers of mice fed with regular (control) or ethanol-containing (alcohol) Lieber-De Carli liquid diets, and of mice fed with diets containing 0.2% (normal) or 2% (overload) carbonyl iron was employed in EMSA. Cold competition with unlabeled Smad consensus (cons.) oligonucleotides and 32P-labeled mutant Smad probe were employed to test the specificity of DNA binding, as described in Materials and Methods. The arrows indicate specific Smad DNA complexes and unbound (free) probes. The results are representative of multiple EMSA experiments ().
