For all fly behavioral studies, groups of 25 adult flies (2–5 days-old, grown in parallel) were collected under brief CO2 anesthesia, transferred to fresh food vials, and housed overnight at 25°C and 55% relative humidity. Each group of 25 flies constituted n=1 (7,425 total flies were used for the reported behavioral studies). Sensitivity and rapid tolerance to ethanol (provided as a vapor) were determined in flies using ethanol Rapid Iterative Negative Geotaxis (eRING) assays at 25°C and 55–65% relative humidity as described (Bhandari et al., 2009). Internal ethanol concentrations in flies exposed to ethanol in eRING assays were determined as described previously (Bhandari et al., 2009). Additional details are provided as Supplementary Material.
