Activation of signaling molecules induced by LPS in vivo is inhibited by ethanol. Mice were treated with ethanol at 6 g/kg (by oral gavage with a 32%, v/v solution). Thirty minutes later, LPS (60 μg/mouse) was administered intravenously. Then at the indicated times after LPS administration, peritoneal macrophages were isolated by gavage using ice cold PBS. Cell pellets were immediately extracted and analyzed for the indicated phosphorylated proteins by western blot. Values shown are means ± SEM (n = 5 mice per group). Values at each time point for LPS-treated and LPS plus ethanol-treated animals were compared by Student's t test and significant differences are indicated by ** (p < 0.01).