Ethanol sedation assays were performed essentially as previously described [5], [46]. Samples of 25–30 male flies were allowed to equilibrate for ∼10 min to humidified air in the booz-o-mat apparatus [4] before starting exposure to a 100∶50 mixture of ethanol vapor∶humidified air (100∶50 E∶A). Ethanol exposure commenced at 0 min of the assay and was continuous thereafter. At 2-min intervals, each tube of flies (8 tubes per assay) was twirled and the number of flies that appeared unable to right themselves was scored by an experimenter blinded to sample identity. Typical assay duration was 30 min.
