Extended Data Fig. 2: Midgut Tk+ EEC activity, Tk peptide levels, and transcripts under different nutritional conditions in mated female flies.
From: Protein-responsive gut hormone tachykinin directs food choice and impacts lifespan
a-b, Assessment of calcium-signaling activity measured by CaLexA in Tk+ EECs (a; GFP normalized to tdTomato) and Tk peptide intensity (b) in the whole midgut of mated females fed standard adult-optimized diet (9% sugar and 8% yeast) or starved on 1% agarose for four hours (two-tailed Mann-Whitney U tests). c-d, Effects on calcium signaling (c) and anti-Tk staining intensity (d) of a 3-hour period of refeeding with 10% sugar alone, 10% yeast alone, or both, following 25-hour starvation (Kruskal-Wallis/Dunn’s). e-f, Midgut Tk transcript levels in mated females: (e) animals were starved for 25 h and then refed for 3 h with 10% sugar or 10% yeast, and (f) animals were starved for 22 h and then refed with sugar+yeast medium for a range of time periods before dissection [two-tailed unpaired Student’s t test (e) and one-way ANOVA with Dunnett’s test (f)]. g, Tk+ EEC activity in dissected midguts, reflected in calcium-LexA-driven Luciferase, measured over a range of yeast-feeding durations following normal feeding on adult diet (one-way ANOVA/Dunnett’s). h, Quantification of GCaMP6s signal (ΔF/F₀) averaged across the R1-R3 Tk+ EECs in sugar- and yeast-fed conditions (two-tailed Mann-Whitney U). i, Anti-Tk staining intensity in R5 EECs of animals fed for 15 h on media containing 10% of either (naturally lipid-free) sugar or lipid-depleted casein (two-tailed Mann-Whitney U). j, Tk transcript levels in dissected midguts of animals fed for 15 h on either sugar, yeast, or the peptide mixture peptone (one-way ANOVA/Dunnett’s). k, Tk transcript levels in midguts of animals fed for 15 h on media containing either sugar, yeast, a mixture of amino acids, lipid-depleted yeast, or lipid-depleted casein (one-way ANOVA/Dunnett’s). Sample sizes (N) and p values are indicated in each plot. In a–b and i, N = EECs. In e–h, j, and k, N = midgut samples. Panels a, b, c, d, h, and i indicate medians and 95%-confidence intervals, and panels e, f, g, j, and k indicate means ± SEM. ns, non-significant (p > 0.05).
