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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. 6 — a, Immunohistochemistry of APCs shows NPFR>GFP reporter expression in APCs of mated females. Scale bars 20 μm. b, Quantification of AKH levels within the APCs of fed and 15-hour-starved mated females with and without NPFR knockdown in these cells; n = 17 fed AKH>, n = 19 fed AKH>NPFRi^TRiP, n = 22 starved AKH>, n = 19 starved AKH>NPFRi^TRiP. Representative images are shown below. Scale bars 20 μm. c, Metabolite levels in fed mated females. TAG n = 9 AKH>, n = 10 AKH>NPFRi^TRiP; glycogen n = 10 AKH>, n = 10 AKH>NPFRi^TRiP. d, Survival during starvation of mated females; n = 96 AKH>, n = 96 AKH>NPFRi^TRiP. e, Immunohistochemistry of guts from mated female NPFR>GFP flies showing expression of NPFR reporter in AstC⁺ cells. Scale bars, 25 μm. f, Quantification of the number of midgut cells per gut that showed detectable AstC staining with and without NPFR knockdown in AstC⁺ EECs of fed mated females; n = 8 AstC^gut> guts, n = 11 AstC^gut>NPFRi^TRiP. g, Midgut AstC transcript levels in fed mated females; each n = 6. h–k, AstC⁺ EEC-cell activity levels (h) with representative images (i) quantified on a per-gut basis (j) and the number of AstC⁺ EEC cells showing detectable GFP (k), measured by calcium-reporter system (AstC>LexA::NFAT::VP16; LexAop-GFP, denoted by AstC> in the figure) with or without NPFR knockdown in the AstC⁺ EECs in the midgut. h, n = 182 AstC> cells, n = 255 AstC>NPFR cells; j, each n = 6 guts; k, each n = 6 guts. Scale bars, 50 μm. l, Survival during starvation of mated females; n = 153 AstC^gut> animals, n = 198 AstC^gut>NPFRi^TRiP. All animals were mated females. Bars represent mean ± s.e.m. Box plots indicate minimum, 25th percentile, median, 75th percentile and maximum values. NS, not ignificant. b,c (left), f,g,j, Two-tailed unpaired Student’s t-test. d,l, Gehan–Breslow–Wilcoxon test. c (right), h,k, Two-tailed unpaired Mann–Whitney U-test.

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