FIG. 5.
H2S attenuates TFAM promoter methylation. (A) There are 10 potential CpG methylation loci in the predicted CpG islands between −210 bp and −30 bp of the mouse TFAM promoter. (B) MS-RE-PCR data show that the CpG islands in the TFAM promoter are methylated in both WT and CSE-KO SMCs. (C) MSP analysis validates that CSE deficiency induces but exogenously applied H2S reduces the methylation ratio in the CpG islands of the TFAM promoter. Both WT and CSE-KO SMCs were incubated with NaHS (30 μM) for 48 h. M, methylation-specific primers; U, unmethylation-specific primers. *p<0.05 versus all other groups; #p<0.05 versus WT-SMCs without NaHS treatment. (D) Higher DNA methylation in CSE-deficient SMCs evidenced by bisulfite genomic sequencing. *p<0.05. The experiments in (B, C) were repeated thrice, and 10 clones were randomly chosen for sequencing in (D).