The intermittent-access drinking protocol was adapted from Rhodes et al. [30]. Ethanol-naive mice were singly housed in single grommet cages in a reverse light-dark cycle room (lights off from 10 a.m. to 10 p.m.) and allowed to acclimate for two weeks prior to the study. Following acclimation, home cage water bottles were replaced with a single bottle of 20% v/v ethanol in water at noon for 4 hours on Monday, Wednesday, and Friday, for a total of 8 ethanol sessions. Bottles were weighed before and after each session and mice were weighed once a week. Baseline water consumption was measured one day before the beginning of ethanol access by weighing a water bottle before and after a 4 hour session. Mice had ad libitum access to water when ethanol was not present. Ethanol consumption (g ethanol/kg mouse/4 hrs) was calculated as the difference between bottle weight before and after drinking sessions. Drinking volumes were corrected for spillage by subtracting weight lost from two control bottles of 20% ethanol placed on empty cages for the duration of the sessions. Data were analyzed for statistical significance by 2-way RM ANOVA.
