Effect of acute ethanol on HCV replication. BB7 replicon cells were treated once with 0, 25, 50, or 100 mM ethanol or 20 U/ml of IFN, and RNA and protein was harvested 72 hours later. A, HCV RNA copy number was determined by quantitative real time RT-PCR. The HCV RNA copy number is reported as copies per 10 ng total cellular RNA. Error bars represent standard deviations. B, HCV protein expression in BB7 cells treated with 0, 25, 50, and 100 mM ethanol, and control Huh7 cells. The positions of Stat1, HCV NS3 and NS5A proteins are indicated. The experiment was repeated twice with similar results.