Mice were exposed to ethanol vapor or room air for 4 days (16 hours of exposure,8 hours of abstinence), followed by 72 hours of room air exposure (one cycle). Cycles were repeated 5 times. Mice were treated (i.p.) with a solution containing 1.6g/kg ethanol (CIE) or saline (air) and 1.0 mmol pyrazole (Sigma Aldrich), an ethanol dehydrogenase inhibitor, 30 minutes prior to inhalation treatment. Blood ethanol concentrations were tested after the first and fourth inhalation exposure to ensure physiologically relevant blood ethanol concentrations (Griffin et al., 2009 (link)) (Supplementary Figure 2). The back-to-back CIE protocol (Figure 1A) was modified (back-to-back CIE+drinking protocol, Figure 1B) to examine CIE-induced changes in ethanol drinking.