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Fig. 8 | Epigenetics & Chromatin

Fig. 8

From: Epigenetic modifier alpha-ketoglutarate modulates aberrant gene body methylation and hydroxymethylation marks in diabetic heart

Fig. 8

DNMT3B knockdown and AKG treatment inhibit DNA methylation/hydroxymethylation in H9c2 cells under hyperglycemic conditions. A Representative images of immunofluorescence staining with DNMT3B antibody in knockdown cells treated with normal glucose (NG) concentration (5.5 mM) and high glucose (HG) concentration (33 mM). B Representative images of immunofluorescence staining with 5mC antibody (Green) and nuclei stained with DAPI (Blue) in both DNMT3B knockdown and AKG-supplemented H9c2 cells in the presence of NG or HG. C, D 5mC- and 5hmC-specific dot blot analysis of gDNA isolated from NG- and HG-treated H9c2 cells in the presence of C AKG treatment and D DNMT3B knockdown. Equal loading of gDNA was assessed by methylene blue (MB) staining. Quantitative analysis of dot blot densitometry and 5mC immunofluorescence intensities are shown in the Additional file 1: Fig. S5. Relative mRNA levels of E TGFBR2 and F TGFBR3 in NG- and HG-treated H9c2 cells with or without AKG by qPCR analysis. Relative mRNA levels of G TGFBR2 and H TGFBR3 in NG- and HG-treated DNMT3B knockdown H9c2 cells by qPCR analysis. mRNA expression was normalized to β-actin, and the results are represented as the mean ± SD for three independent experiments. Statistical significance was determined by one-way ANOVA with Tukey’s multiple comparisons post-test, where *p < 0.05, **p < 0.001, ****p < 0.0001 vs NG (5.5 mM) treatment, #p < 0.05, ##p < 0.001 ####p < 0.0001 vs HG (33 mM) treatment

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