Effect of acute ethanol on the Jak-Stat pathway. A, high physiological doses of ethanol activate the ISRE. Huh7 cells were transfected with 0.7 μg of pISRE-luc, and 24 hours later, cells were stimulated with ethanol at the indicated concentrations. Protein lysates were assayed for luciferase activity 6 hours later. Error bars represent standard deviations. The experiment was repeated 6 times with similar results. B, acute ethanol induces Stat1 serine phosphorylation. Huh7 cells were left as untreated controls (C) or treated with 1,000 U/ml of IFN-α (IFN), or 100 mM ethanol (EtOH). Twenty minutes later, equal amounts of whole cell protein extracts were separated by SDS-PAGE, and blotted for phosphorylated Stat1 S727 (toppanel), Stat1 Y701 (second panel), Stat2 Y690 (third panel), and total forms of the Stat1 protein (lower panel). The figure is representative of 3 independent experiments, which yielded similar results.