Extended Data Fig. 7: Expression of Dh31R and Crz in brain Crz+ neurons is required for the aphrodisiac effect of amino acids.
From: A nutrient-specific gut hormone arbitrates between courtship and feeding

(a) Validation of Dh31R knockdown. qRT-PCR results show that Dh31R expression level is reduced in flies containing the Actin-GAL4 and UAS-Dh31R-RNAi transgenes. RNA was extracted from 5 male flies per experiment, with 6 experiments for each genotype. One-way ANOVA followed by Tukey’s test. (b–d) Schematic indicates genetic strategies to perturb Crz+ neurons in the brain and VNC (b), only in the brain (c), and only in the VNC (d). Representative confocal images from 3 samples show the different intersections between Crz-GAL4 and otd-nls::flp (corresponding genotypes are shown below the images). Samples were immuostained with anti-GFP (green), nc82 (blue). Scale bar, 50 μm. The courtship effect of Dh31R knockdown in both brain and VNC, in brain only or in VNC only. Each data point represents one experiment. For each condition, the total number of matches (n) is indicated in the figure. p-value was determined by Chi-square test to indicate whether males of a given genetic manipulation respond to amino acids. Blue bar indicates average copulation percentage, and the dashed line indicates chance level. AAs, amino acids (200 mM). S, sucrose (50 mM). (e) Validation of Crz knockdown. Immunostaining of anti-Crz (red) showed Crz is undetectable in brain Crz+ neurons in Crz-Gal4, UAS-Crz-RNAi flies, based on 3 independent experiments. Scale bar, 50 µm. (f–g) The courtship effect of Crz knockdown in brain Crz+ neurons (f) or in VNC Crz+ neurons (g). For each condition, the total number of matches (n) from 5 independent experiments is shown in the figure. p-value was determined by Chi-square test to indicate whether males of a given genetic manipulation respond to amino acids. Blue bar indicates average copulation percentage, and the dashed line indicates chance level. AAs, amino acids (200 mM). S, sucrose (50 mM).