Extended Data Fig. 7: KD and Na-β-OHB treatment increase the levels of K108bhb and K147bhb.
From: Ketogenic diet reshapes cancer metabolism through lysine β-hydroxybutyrylation
a Conservation of Aldob kbhb sites in multiple organisms. b Specificity of Aldob K108bhb antibody determined by a dot blot assay. Synthetic peptides with K108bhb, K108Ac or unmodified K108 were blotted with the Aldob K108bhb antibody. c Specificity of Aldob K147bhb antibody determined by a dot blot assay. Synthetic peptides with K147bhb, K147Ac or unmodified K147 were blotted with the Aldob K147bhb antibody. d, e Immunoblotting shows the Aldob K108bhb level in male mice livers (n = 3 per group). The mice were fed with a control diet or KD for 5 weeks. The graph showed ratios of K108bhb to Tubulin quantified by densitometry using Image J software. p value: 1.3e − 5. f, g Immunoblotting shows the Aldob K147bhb level in male mice livers (n = 3 per group). The mice were fed with a control diet or KD for 5 weeks. The graph showed ratios of K147bhb to Tubulin quantified by densitometry using Image J software. h, i HCC-LM3 cells were treated with 10 mM Na-β-OHB for 24 h, and whole cell lysates were probed by immunoblotting with anti-K108bhb or anti-Tubulin antibodies. The graph showed ratios of K108bhb to Tubulin quantified by densitometry using Image J software. j, k HCC-LM3 cells were treated with 10 mM Na-β-OHB for 24 h, and whole cell lysates were probed by immunoblotting with anti-K147bhb or anti-Tubulin antibodies. The graph showed ratios of K147bhb to Tubulin quantified by densitometry using Image J software. Experiments in (h, j) were performed in triplicate, each dot represents an independent experiment with three technical replicates (i, k). Statistical data are presented as mean ± SEM (e, g, i, k), P values were determined by unpaired two-tailed t test (e, g, i, k).
