The amount of dephosphorylated Y1176 of L1 is decreased by ethanol in vivo. (A) Animals were suckled (SC) or intubated and treated with 0 (IC), 4.5, 5.25 or 6.0 g/kg of ethanol. Shown is a male:female pair in each group. Cerebella were harvested 2 hours after the second alcohol feeding and lyzed. Equal amounts of protein from each supernatant were run on SDS gel electrophoresis and immunoblotted for dephosphorylated Y1176-L1 using 74-5H7, a monoclonal antibody that specifically recognizes unphosphorylated Y1176 in the cytoplasmic domain of L1. Blots were stripped and reblotted for total L1 using a polyclonal antibody against the cytoplasmic domain of L1 as a loading control. (B) Densitometric quantification of dephosphorylated Y1176 corrected for total L1 is plotted for each treatment group. The values of 3 different experiments are shown. The bar indicates the mean of the values +/- S.D. All animals which received ethanol had significantly less dephosphorylated L1 than either the SC or the IC controls. ANOVA single factor, p<10-7; *vs both IC and SC, p< 0.05, paired t-test.