Skip to main content
. 2019 Jan 24;9:742. doi: 10.1038/s41598-018-36941-9

Figure 1.

Figure 1

Effects of NaBut and β-hydroxybutyrate molecules on histone acetylation in multiple cell types. (A) HEK293 cells were incubated for 18 hours with 5 mM NaBut or increasing concentrations of NaR-βOHB (10–40 mM range). (B) HEK293 cells were incubated for 18 hours with NaBut, the racemic mix of β-hydroxybutyrate sodium salt (Na R/S-βOHB), R-β-hydroxybutyric acid (R-βOHB) and S-β-hydroxybutyric acid (S-βOHB) at the indicated concentrations. Separation bars indicate non-contiguous lanes on the same image acquisition (the original blots are shown in the supplementary information file). (C) HMEC-1 were incubated for 8 or 18 hours with SAHA, NaBut or NaR-βOHB at the indicated concentrations. (D) L6 myotubes were incubated with 10 mM NaBut, 40 mM R-βOHB or 40 mM S-βOHB for the indicated times. (A–D) Acid-extracted histones were immunoblotted with antibodies anti H3K9/14Ac, anti-acetyl-H3K9Ac, anti-acetyl-H3 (H3KAc), anti-acetyl lysine, anti total H3 or stained with coomassie blue as indicated. (A,B) quantification of acetyl immunoblotting signals normalized to total histone H3 content. n = 3. Error bars represent means ± SEM from 3 independent experiments. (*p < 0.05, versus control group (−) using one-way ANOVA). Quantifications for panels A (H3K9/14Ac and Acetyl lysine blots), C, D are shown in Supplementary Fig. 1.