We previously reported that genes in the Chloride Intracellular Channel (CLIC) family influence ethanol sensitivity in flies, worms and mice (Bhandari et al., 2012). In the previous Drosophila experiments, we used eRING assays to measure ethanol sensitivity in flies harboring two independent Clic transposon insertions marked with mini-w. Given the data in Figure 1, it seemed possible that the decreased ethanol sensitivity exhibited by Clic mutants—as measured in eRING assays—could be confounded by the presence of the mini-w marker. We therefore used ethanol sedation assays to re-examine ethanol sensitivity in the same two Clic transposon mutants. Consistent with our previous eRING studies, we found that both Clic mutants had decreased ethanol sensitivity in ethanol sedation assays (Figure 7A and B). Similarly, a 40±4% knockdown (one-sample t test, p=0.0007, n=5) of Clic via ubiquitous da-Gal4-driven expression of the Clic RNAi transgene v105975 also lessened ethanol sensitivity in sedation assays without having a major effect on internal ethanol concentrations (Figure 7C and E). Nervous system expression of Clic RNAi v105975 also decreased ethanol sensitivity without substantively impacting internal ethanol concentrations (Figure 7D
