Fig. 7: Mitochondrial protein β-hydroxybutyrylation and acetylation may be responsible for the regulation of mitochondrial maturation by ketone body β-HB. | Cell Discovery

Fig. 7: Mitochondrial protein β-hydroxybutyrylation and acetylation may be responsible for the regulation of mitochondrial maturation by ketone body β-HB.

From: Neonatal ketone body elevation regulates postnatal heart development by promoting cardiomyocyte mitochondrial maturation and metabolic reprogramming

Fig. 7

a Western blot detection of mitochondrial protein K-ac in the hearts of WT mice at E18.5d, P1d and P7d. b Western blot detection of mitochondrial protein K-bhb in the hearts of WT mice at E18.5d, P1d and P7d. c Western blot detection of mitochondrial protein K-ac in the hearts of WT and KO mice at P5d. d Western blot detection of mitochondrial protein K-bhb in the hearts of WT and KO mice at P5d. e Levels of Ac-CoA in the hearts of WT and KO mice at P5d (WT, n = 6; KO, n = 4). f Western blot detection of protein K-ac and K-bhb of mitochondria isolated from KO hearts at P5d treated with ddH2O or β-HB. g Western blot detection of protein K-ac and K-bhb of mitochondria isolated from WT hearts at P5d treated with ddH2O or Ac-CoA. h Immunoprecipitation assays on WT and KO hearts at P5d. IP: Anti-K-hbb. IB: IDH2, SDHA, MDH2. i Immunoprecipitation assays on WT and KO hearts at P5d. IP: Anti-K-ac. IB: IDH2, SDHA, MDH2. j Relative mitochondrial IDH (mIDH) activity of mitochondria isolated from WT and KO hearts at P5d treated with ddH2O or 5 mM β-HB (WT, n = 4; KO, n = 4; KO + β-HB, n = 5). k Relative SDH activity of mitochondria isolated from WT and KO hearts at P5d treated with ddH2O or 5 mM β-HB (n = 3 for each group). l Relative MDH activity of mitochondria isolated from WT and KO hearts at P5d treated with ddH2O or 5 mM β-HB (n = 3 for each group). m The OCR of isolated mitochondria in KO mouse hearts at P5d treated with ddH2O or 5 mM β-HB was detected by an extracellular metabolic flux analyzer (n = 3 for each group). n Relative mIDH activity of mitochondria isolated from 293T cell lines transfected with WT-IDH2 or IDH2-K179R (n = 3 for each group). o Relative SDH activity of 293T cell lines transfected with WT-SDHA or SDHA-K263R (n = 5 for each group). p The OCR of 293T cell lines transfected with WT-IDH2 or IDH2-K179R (n = 6 for each group). q The OCR of 293T cell lines transfected with WT-SDHA or SDHA-K263R (WT-SDHA, n = 5; SDHA-K263R, n = 6). r Western blot detection of pHH3, total histone H3 (T-H3) and PCNA in 293T cell lines transfected with WT-IDH2, IDH2-K179R, WT-SDHA or SDHA-K263R. Data are presented as means ± SEM. and the P values were determined by unpaired two-tailed Student’s t-test. *P < 0.05; **P < 0.01; ***P < 0.001.

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