Extended Data Fig. 3: Nutrient-intake regulation, metabolic consequences, and sleep effects of TkR99D deficiency in the APCs and loss of AKH signaling in mated female flies.
From: Protein-responsive gut hormone tachykinin directs food choice and impacts lifespan
a-d, Analysis of midgut Tk peptide and transcript (a, b, d), and slif (c) transcript levels in flies expressing Tk+-EEC-specific RNAi against slif (a, c), a range of other amino-acid transporters (b), or the central regulatory gene Gcn2 (d) in animals fed 15 h with yeast. (two-tailed unpaired Student’s t tests in a, c, d; one-way ANOVA/Dunnett’s in b.) e, Calcium activity (CaLexA>GFP normalized to tdTomato) in all Tk+ midgut EECs of animals fed for 3 h on normal adult optimized diet (ND) or on medium containing hydrogen peroxide (two-tailed Mann-Whitney U). f, anti-Tk staining intensity of R5 EECs in 15 h sugar-fed controls and animals expressing RNAi against TrpA1 in Tk+ EECs (two-tailed Mann-Whitney U). g, 1-hour FlyPAD assay of sugar-directed feeding behavior for animals expressing RNAi against slif in the Tk+ EECs (two-tailed Mann-Whitney U). h-i, 1-hour dye-feeding assay for sugar consumption for UAS-slif-RNAi alone (two-tailed Mann-Whitney U tests). j-k, FlyPAD feeding behavior (j) and dye-feeding consumption (k) of yeast in controls and animals expressing RNAi against slif in the Tk+ EECs [two-tailed Mann-Whitney U (j) and Kruskal-Wallis/Dunn’s (k)]. l, Dye-feeding consumption of yeast in animals exhibiting attenuated TOR signaling in the Tk+ EECs (Kruskal-Wallis/Dunn’s). m-n, One-hour dye-feeding assay for yeast intake for UAS-slif-RNAi alone (two-tailed Mann-Whitney U tests). Behavioral experiments were performed following 15 h of yeast feeding. Sample sizes (N) and p values are indicated in each plot. In a, c, and d, N = midgut samples. In b, e, and f, N = EECs. In g–n, N = animals per group. Box plots show the median, the bounds of the box (25th and 75th percentiles), and whiskers representing the minimum and maximum values. Dot plots in a-d include mean ± SEM, and in e, f, g, and j indicate the median and 95%-confidence interval. ns, non-significant (p > 0.05).
