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. 2021 Nov 25;12:6838. doi: 10.1038/s41467-021-27141-7

Fig. 9. DNMT1 silences Myod1 expression via interacting with PRDM16.

Fig. 9

A Comparison of genome-wide alterations in chromatin accessibility landscape assessed by ATAC-seq with the corresponding gene expression assessed by RNA-seq in iBAT of UTXKO and fl/Y mice fed HFD for 12 weeks (n = 3 replicates per group). B ATAC-seq analysis of chromatin accessibility at Prdm16 gene locus in iBAT of UTXKO and fl/Y mice fed HFD for 12 weeks (n = 3 replicates per group). C Quantitative RT-PCR analysis of Prdm16 mRNA in iBAT of LFD- or HFD-fed mice (n = 8/Group). *Indicates statistical significance between the two groups by two-tailed unpaired Student’s t-test. D, E ChIP assay of UTX binding to Prdm16 promoter (D, n = 4/group) and ChIP assay of H3K27me3 levels at Prdm16 promoter (E, n = 4/group) in iBAT of LFD- or HFD-fed mice. *Indicates statistical significance between the two groups by Mann–Whitney’s nonparametric U test in (D) and two-tailed unpaired Student’s t-test in (E). F ChIP assay of H3K27me3 levels at Prdm16 promoter in control or Utx knockdown BAT1 brown adipocytes treated with isoproterenol (n = 4/Group). *Indicates statistical significance between the two groups by Mann–Whitney’s nonparametric U test. G–H Pyrosequencing analysis of DNA methylation at Myod1 promoter (G, n = 6/group) and Myod1 expression (H, n = 8/group) in BAT1 brown adipocytes transfected with scramble or Utx siRNA. *Indicates statistical significance between the two groups by Mann–Whitney’s nonparametric U test in (G) and two-tailed unpaired Student’s t-test in (H). I, J Pyrosequencing analysis of DNA methylation at Myod1 promoter (I, n = 4/group) and Myod1 expression (J, n = 8 for Scramble and 7 for Prdm16 siRNA) in BAT1 brown adipocytes transfected with scramble or Prdm16 siRNA. *Indicates statistical significance between the two groups by Mann–Whitney’s nonparametric U test in (I) and two-tailed unpaired Student’s t-test in (J). K Pyrosequencing analysis of DNA methylation at Myod1 promoter in BAT1 brown adipocytes transfected with pSPORT6 or pSPORT6 encoding Prdm16 overexpressing plasmids (n = 4/group). *indicates statistical significance between the two groups by Mann–Whitney’s nonparametric U test. All data are expressed as mean ± SEM.