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. 2021 Sep 13;12(9):848. doi: 10.1038/s41419-021-04111-x

Fig. 5. KAT7 reversed the upregulation effect of miR-134-5p on intracellular ROS accumulation by promoting the transcriptions of MnSOD and catalase.

Fig. 5

A Bioinformatics databases TargetScan and miRDB forecasted there were 19 common target genes of miR-134-5p. The (B) mRNA and (C) protein levels of lysine acetyltransferase 7 (KAT7) in H9C2 cells and primary rat cardiac fibroblasts treated with EAT-CM or EAT-CM + miR-134-5p inhibitor were measured by qRT-PCR and western blot, respectively. β-actin was used as an internal control. D–F H9C2 cells and primary rat cardiac fibroblasts were divided into four groups: miR-134-5p inhibitor, the negative control of miR-134-5p inhibitor (inhibitor NC), miR-134-5p inhibitor + si- lysine acetyltransferase 7 (KAT7)-1, miR-134 -5p inhibitor + si-KAT7-2. All the cells were cultured in EAT-CM. D The intracellular ROS level was analyzed by flow cytometry using a DCFH-DA fluorescence probe. The (E) mRNA and F protein levels of manganese superoxide dismutase (MnSOD) and catalase. G H9C2 cells and primary rat cardiac fibroblasts were divided into three groups: EAT-CM, EAT-CM + miR-134-5p inhibitor, EAT-CM + miR-134-5p inhibitor + si-KAT7. Histone H3K14 acetylation (H3K14Ac) level of cells was analyzed using Western blot. H3 was used as an internal control. H–K H9C2 and primary rat cardiac fibroblasts were divided into EAT-CM and EAT-CM + miR-134-5p inhibitor groups. H, J The H3K14Ac levels of 2 kb, 1 kb, and 0.1 kb upstream of transcription start site (TSS) of MnSOD/catalase gene were measured by chromatin immunoprecipitation (CHIP)-PCR. I, K The combination between KAT7 and 2 kb, 1 kb, and 0.1 kb upstream of TSS of MnSOD/ catalase gene was measured by CHIP-PCR. L, M H9C2 cells and primary rat cardiac fibroblasts were divided into three groups: EAT-CM, EAT-CM + miR-134-5p inhibitor, EAT-CM + miR-134-5p inhibitor + si-KAT7. The H3K14Ac levels of 1 kb upstream of TSS of MnSOD gene and 0.1 kb upstream of TSS of catalase gene were measured by CHIP-PCR. B, C, H–K *P < 0.05, **P < 0.01 vs EAT-CM. D–F **P < 0.01 vs inhibitor NC; #P < 0.05, ##P < 0.01 vs miR-134-5p inhibitor. G **P < 0.01 vs EAT-CM; ##P < 0.01 vs EAT-CM + miR-134-5p inhibitor.