Downregulation of circ-ZNF609 Promotes Heart Repair by Modulating RNA N6-Methyladenosine-Modified Yap Expression
circ-ZNF609 is upregulated during myocardial I/R injury. (a) The expression of circ-ZNF609 was validated by RT-PCR followed by Sanger sequencing. mmu: total RNA extracted from mouse heart; rat: total RNA extracted from rat heart; hsa: total RNA extracted from AC16 cardiomyocyte cell line. (b) qRT-PCR for the abundance of circ-ZNF609 and Zfp609 mRNA in mouse hearts treated with RNase R (. wells/group). (c) Expression of circ-ZNF609 in NRCMs (neonatal rat cardiomyocytes) compared to NRCFs (neonatal rat cardiac fibroblasts) ( wells/group). (d) RNA fluorescence in situ hybridization assay (FISH) of circ-ZNF609 in NRCM (scale μm). (e) qRT-PCR indicates the abundance of circ-ZNF609 in the cytoplasm or nucleus ( wells/group). (f) circ-ZNF609 localized in the cytoplasm of cardiomyocytes, as evidenced by MS2-tagged circ-ZNF609 cotransfection with MS2-GFP-NLS (scale μm). (g) Increased circ-ZNF609 in murine hearts from 4 weeks post-I/R injury versus sham control ( mice/group). (h) The expression of circ-ZNF609 was increased in OGD/R-induced NRCM apoptosis model ( wells, /group). (i) Top: the schematic diagram of definition of cardiac infarct, border, and remote zones. Bottom: the expression level of circ-ZNF609 was analyzed by qRT-PCR in the infarct, border, and remote zones of mouse acute I/R hearts compared to the sham group (, ns: nonstatistically significant. mice/group). NRCM: neonatal rat cardiomyocyte; I/R: ischemia/reperfusion; OGD/R: oxygen-glucose deprivation/reperfusion. Data are presented as (b, c, e, g–i) Independent-sample -test.