Ischemic Postconditioning Protects against Aged Myocardial Ischemia/Reperfusion Injury by Transcriptional and Epigenetic Regulation of miR-181a-2-3p

Figure 5

Cooperation of DNMT3b and HDAC2 involved in the regulation of miR-181a-2-3p and autophagy in aged cardiomyocytes. After the aged cardiomyocytes were transfected with sh-DNMT3b and sh-HDAC2 in HPostC, then (a) MassARRAY analysis detected the DNA methylation level of miR-181a-2-3p promoter. (b) ChIP measured enrichment of H3K14ac at miR-181a-2-3p promoter (). (c) qRT-PCR was performed to measure miR-181a-2-3p expression (). (d–f) The levels of DNA methylation and H3K14ac at miR-181a-2-3p promoter and the expression of miR-181a-2-3p were detected after the aged cardiomyocytes were treated with NA (DNMT3b inhibitor) and romidepsin (Rdps, HDAC2 inhibitor) (). (g) Western blot analysis of LC3B-II and p62 expression in aged cardiomyocytes transfected with sh-DNMT3b and/or sh-HDAC2 in the presence of CQ (). (h) Representative confocal fluorescent images of mRFP-GFP-LC3 after aged cardiomyocytes were treated with NA and Rdps (scale  μm). Numbers of autophagosomes and autolysosomes in each cell were quantified (). Data were presented as the .

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