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. 2023 Sep 25;14:5971. doi: 10.1038/s41467-023-41570-6

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© The Author(s) 2023

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. 2 — a Energy expenditure (n = 19:27:13; repeated measures analysis of variance [ANOVA] P < 0.0001), b accumulative food intake for three weeks (n = 8:24:10; repeated measures ANOVA P = 0.70), c wheel rotations (n = 9:13:5; P-for-trend = 0.55), d distance traveled (n = 5:20:7 P-for-trend = 0.27), and e behaviors monitored by HomeCage systems (n = 5:20:7; P-for-trend = 0.88, 0.54, 0.48, 0.12, 0.31, 0.92, 0.65, 0.19, and 0.52 respectively) of the Aldh2 homozygous knock-in (KI), heterozygous KI (HE), and wild-type (WT) mice. f Diet-induced thermogenesis measured by rectal temperature after high-fat high-sucrose feeding (repeated measures ANOVA P < 0.0001) and g cold-induced thermogenesis measured by rectal temperature of mice in 4 °C condition (n = 9:24:10; repeated measures ANOVA P = 0.29). h Isoproterenol-induced energy expenditure (n = 4:8:4) of Aldh2 homozygous KI, HE, and WT mice. The arrow head indicates the injection time (repeated measures ANOVA P = 0.15). i Glycemic levels during the insulin sensitivity test (repeated measures ANOVA P = 0.055) and j intraperitoneal glucose tolerance tests (repeated measures ANOVA P = 0.0049), and k glycemic levels during the oral glucose tolerance test of Aldh2 KI, HE, and WT mice (n = 49:24:42; repeated measures ANOVA P = 0.053). l Insulin levels following oral glucose load (n = 35:14:44; repeated measures ANOVA P = 0.0048 in duplicates). Fasting serum m leptin (n = 9:10:11; P-for-trend = 0.0002 in duplicates), n free fatty acid, and (n = 10:10:10; P-for-trend = 0.0008 in duplicates), o adiponectin (n = 10:10:10; P-for-trend = 0.0008 in duplicates), and p ethanol (n = 10:10:10; P-for-trend = 0.0032 in duplicates) concentration of Aldh2 homozygous KI, HE, and WT mice on high-fat high-sucrose diet. Figure (a) was analyzed using the generalized linear model. Figures (a, b, f–l) were further analyzed using repeated measures ANOVA. Figures (b–p) were analyzed using tests for linear trend. All data are presented as mean and standard error (S.E.M.). The n values represent biological repeats and the number of technical repeats were expressed as duplicates or triplicates. The asterisks indicate two-sided *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.001.