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. 2021 Jul 13;12:4285. doi: 10.1038/s41467-021-24505-x

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© The Author(s) 2021

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. 3 — a O₂ consumption and CO₂ production, measured with indirect calorimetry. b CO₂ production measured in Nf1^P1 mutants and w^CS10 controls, plotted in 1-h bins across the circadian photoperiod. c Quantification of CO₂ production from (b) during day and night periods. ***p < 0.001 (Sidak, two-sided; n = 14 per genotype). d O₂ consumption measured in Nf1^P1 mutants and w^CS10 controls. e Quantification of O₂ consumption from (d) during day and night periods. Day experimental ***p < 0.001 re: control, night experimental **p = 0.003 re: control. (Sidak, two-sided; day: n = 14 per genotype, night: n = 13 control, n = 14 Nf1^P1). f Respirometry quotient in Nf1^P1 mutants and w^CS10 controls. ***p < 0.001 (Mann–Whitney, two-sided; n = 14). g CO₂ production, comparing pan-neuronal nSyb > RNAi with heterozygous controls. h Quantification of CO₂ production from (g) during day and night periods. Day experimental ***p = 0.001 re: UAS-Nf1 RNAi/+, **p = 0.001 re: nSyb-Gal4/+. Night experimental **p = 0.004 re: UAS-Nf1 RNAi/+, **p = 0.002 re: nSyb-Gal4/+. (Sidak, two-sided; n = 6 UAS-Nf1 RNAi/+, n = 7 nSyb-Gal4/+, n = 6 experimental group). i O₂ consumption, comparing pan-neuronal nSyb > RNAi lines with heterozygous controls. j Quantification of O₂ production from (i) during day and night periods. Day experimental ***p = 0.0001 re: UAS-Nf1 RNAi/+, ***p = 0.003 re: nSyb-Gal4/+. Night experimental **p = 0.003 re: UAS-Nf1 RNAi/+, ***p = 0.001 re: nSyb-Gal4/+. (Sidak, two-sided; n = 6 UAS-Nf1 RNAi/+, n = 7 nSyb-Gal4/+, n = 6 experimental group). k Respirometry quotient in nSyb > RNAi lines and heterozygous controls. Experimental *p = 0.014 re: UAS-Nf1 RNAi, *p = 0.011 re: nSyb-Gal4/+ (Sidak, two-sided; n = 6 UAS-Nf1 RNAi/+, n = 7 nSyb-Gal4/+, n = 6 experimental group). Each “n” represents a single respirometer containing 25 animals. Box plots—box: 1st to 3rd quartiles, median: line, whiskers: min–max, individual data points: circles. Data are presented as mean values ± SEM in (b, d, g, and i).