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. 2023 Sep 27;622(7983):619–626. doi: 10.1038/s41586-023-06585-5

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© The Author(s) 2023, corrected publication 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a, Metabolic flux assay of Ctrl^Cre and Cpt1b^cKO hearts perfused with [¹³C]palmitate (n = 5 each). b, Volcano plot showing different metabolites in Ctrl^Cre and Cpt1b^cKO hearts (n = 8 each). Symbol colors indicate increased (red), decreased (blue) or unchanged (black) metabolites in Cpt1b^cKO CMs. Dashed horizontal line indicates the threshold of changed metabolites with a P value of < 0.05; dashed vertical lines indicate a fold change of >2 or <0.5. c, Quantification of metabolites associated with the Krebs cycle and glycolysis, as well as amino acids, in Cpt1b^cKO hearts (pyruvate, n = 6 each; all other metabolites, n = 8 each). Dashed outline indicates the change in αKG. d,e, Quantification of BCAA catabolism-associated metabolites (n = 7 for d; n = 8 for e). KIV, α-ketoisovalerate; KMV, α-keto-β-methylvalerate. f,g, Metabolic flux assay of Ctrl^Cre and Cpt1b^cKO hearts perfused with [¹³C]glucose (f) or [¹³C]isoleucine (g) (n = 6 each). h, Western blot analysis of enzymes involved in αKG generation and catabolism (n = 3 each). Pan-actin was used as a loading control. For gel source data, see Supplementary Information. i, Enzymatic activity of OGDH in CMs from adult Ctrl^Cre and Cpt1b^cKO mice (n = 3 each). j, Comparison of Krebs cycle metabolites in CMs from control hearts (n = 6) and Ogdh^iKO hearts (n = 5). Dashed outline indicates the change in αKG k, Trichrome staining of heart sections from control and Ogdh^iKO mice, 3–4 weeks after termination of TAM treatment for Ogdh deletion. RV, right ventricle; LV, left ventricle. Scale bars, 500 μm. l, Quantification of CMs in adult control and Ogdh^iKO hearts (n = 3 each). m–o, Quantification of Ki67⁺ (m), pH3⁺ (n), aurora B⁺ (o) and sarc-actinin⁺ CMs on heart sections from control and Ogdh^iKO mice (m, control n = 5; Ogdh^iKO n = 4; n and o, n = 4 each). Error bar represents mean ± s.e.m. n numbers refer to individual mice. Two-tailed, unpaired Student t-tests were used for statistical analysis in a,c–g,i,j,l–o. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

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