Extended Data Fig. 1: Compensatory appetite for EAAs in amino-acid-deprived male flies, virgin female flies, poxn-mutant flies and mated female flies in which Ir76b⁺ neurons or Dh44⁺ neurons were inactivated.

a–c, Two-choice preferences of wild-type w¹¹¹⁸ mated female (a), male (b) and virgin female (c) flies that had been fed (−) or had been deprived of amino acids for the indicated durations (5, 24, 48 and 72 h). Flies were given a choice between: l-EAA + l-NEAA + d-glucose and d-EAA + d-NEAA + d-glucose (a) or l-EAA + d-glucose and d-EAA + d-glucose (b, c). d, Food preferences of w¹¹¹⁸ mated female flies fed with diets containing different amounts of dietary yeast were given a choice between l-EAA + d-glucose and d-EAA + d-glucose. e, Food preferences of flies in which Ir76b⁺ neurons had been inactivated by expressing tetanus toxin (TNT) (Ir76b-LexA>LexAop-TNT flies) were fed (−) or were deprived of amino acids for 48 h before the two-choice assay: l-EAA + d-glucose versus d-EAA + d-glucose. Flies carrying Ir76b-LexA alone or LexAop-TNT alone were used as controls. f, Two-choice preferences of control w¹¹¹⁸ and poxn^−/− flies that had been deprived of amino acids for 48 h. g, Two-choice preferences of flies carrying Dh44-Gal4 and UAS-Kir2.1 that had been fed (−) or had been deprived of amino acids for 48 h. Flies carrying Dh44-Gal4 alone or UAS-Kir2.1 alone were used as controls. Inactivation of Dh44⁺ neurons was achieved by expressing inwardly rectifying potassium channels (Kir2.1) in Dh44⁺ neurons. Data are mean ± s.e.m. P values are indicated; unpaired two-tailed t-test in a; one-way ANOVA with Tukey’s post-hoc test in b–g. n is experimental trials. Sample sizes and statistical analyses are shown in Supplementary Table 1.

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