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. 2022 Nov 7;4(11):1532–1550. doi: 10.1038/s42255-022-00672-z

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 8 — a, Sugar intake by dye assay. n = 8 w¹¹¹⁸, n = 9 AKH^−/−. b, Consumption preference, n = 17 AKH>, n = 20 AKH>AKHi^KK, n = 20 AKH^ts>, n = 28 AKH^ts>AKHi^KK. c, Heat map of 30 min tracking of 12 females per genotype. Ratio of time spent on yeast versus sugar patches, n = 3. d,e, Sugar and yeast intake using dye assay, at 29 °C for TrpA1 activation. d, n = 12 AKH>, n = 14 AKH>TrpA1, n = 15 AKH>TrpA1+AKH, n = 15 AKH>TrpA1+AKHi. e, n = 15 AKH>, n = 8 AKH>TrpA1, n = 7 AKH>TrpA1+AKH, n = 15 AKH>TrpA1+AKHi. f, Yeast intake measured by CAFÉ assay. n = 6 AKH>, n = 15 AKH>AKHi^KK, n = 17 w¹¹¹⁸, n = 16 AKH^−/−. g, APCs staining and cell activity, left two panels show n = APCs from 18 w¹¹¹⁸ virgins, n = 18 w¹¹¹⁸ females mated to SP-deficient males (SP⁰/Df(3L)delta130), n = 16 w¹¹¹⁸ females mated to SP⁺ males. Right two panels show n = 15 w¹¹¹⁸ virgins without NPF injection, n = 13 w¹¹¹⁸ virgins with NPF injection. h, AKH staining intensity in mated females with SPR knockdown in the ppk⁺ neurons with or without NPF injection, n = 17 ppk>SPRi mated females, n = 13 ppk>SPRi mated females with NPF injection. i, Yeast consumption measured by dye assay, n = 18 w¹¹¹⁸ virgins, n = 16 AKH^−/− virgins, n = 10 mated w¹¹¹⁸ females, n = 16 mated AKH^−/− females. j, Consumption preference using the two-choice dye assay; n = 17 w¹¹¹⁸ virgins, n = 24 AKH^−/− virgins, n = 19 mated w¹¹¹⁸ females, n = 22 mated AKH^−/− females. k, Cumulative behavioural preference using flyPAD. Lines represent means, and shading indicates s.e.m., n = 21 w¹¹¹⁸ virgins, n = 22 AKH^−/− virgins, n = 23 mated w¹¹¹⁸ females, n = 20 mated AKH^−/− females. l, Preference measured using the two-choice dye assay at 29 °C for TrpA1 activation, n = 23 AKH> virgins, n = 42 AKH>, n = 22 AKH>TrpA1, n = 24 AKH>AKHi mated females. m, A model of the NPF-AKH axis. Bars represent mean ± s.e.m. Box plots indicate minimum, 25th percentile, media, 75th percentile and maximum. Inj., injection; NS, not significant. a,b (right), c,f, Two-tailed unpaired Student’s t-test. b (left), g (right), h, Two-tailed unpaired Mann–Whitney U-test. d,e,g (left), i,k, One-way ANOVA with Tukey’s post hoc test. j, One-way ANOVA with Dunnett’s multiple comparison test or Kruskal–Wallis with Dunn’s multiple-comparisons test. l, One-way Kruskal–Wallis ANOVA with Dunn’s multiple-comparisons test.

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