Acute alcohol inhibits the antiviral actions of IFN. A, Huh7 cells were left untreated (Ctrl), or treated with 1,000 U/ml of IFN-α alone (IFN) or with IFN-α plus 100 mM ethanol (IFN+EtOH). Proteins were probed for Stat1 Y701 (top panel), and total Stat1 proteins (lower panel). B, BB7 replicon cells were treated with or without 100 mM ethanol, followed immediately by 0, 0.1, 1, 10, 20, 100 IU/ml of IFN-α, and whole cell protein extracts were harvested 48 hours later. Equal amounts of total cellular protein were analyzed for the presence of Stat1, HCV NS5A and NS3 proteins, and p42/44 MAPK by western blot analysis. C, quantitation of HCV protein expression shown in panel B. For each lane, pixel intensities of NS3 and NS5A bands were normalized to the total p42/44 pixel intensity, and the fold decrease relative to untreated cells was calculated. The figure is representative of 2 independent experiments that produced identical results.