Flies were tested on the Fly Liquid-Food Interaction Counter (FLIC) system as previously described to monitor ethanol feeding (Ro et al., 2014) after exposure to a choice or fixed diet for 10 days. We filled the liquid-food reservoir with 15% (v/v) ethanol + 5% (w/v) sucrose in 4 mg/l MgCl2. We synchronized the feeding state prior to loading flies onto the FLIC system. Flies were removed from their food environment to kimwipes with 2 ml of water 2 hr before their natural breakfast time. At the peak of their normal breakfast meal (lights on), we flipped flies back onto their respective choice or fixed diets and let them feed for 2 hr. Following breakfast feeding, flies were briefly fasted for 3 hr on a wet kimwipe with 2 ml of water. We began the FLIC recording software before aspirating flies into the system and analyzed food or ethanol interactions for 6 hr. Flies were anesthetized briefly on ice and manually aspirated into the Drosophila feeding monitors (DFMs). Each DFM was loaded with flies from at least two treatment groups to reduce technical bias from the DFM signals. FLIC data were analyzed using custom R code, which is available at wikiflic.com. Default thresholds were used for analysis except for the following: minimum feeding threshold = 20, minimum events = 1, and tasting threshold = (10, 20). Flies that had zero feeding events over the testing interval were removed from the analysis.

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