MiR-486 regulates cardiomyocyte apoptosis by p53-mediated BCL-2 associated mitochondrial apoptotic pathway

Fig. 1. Identification of cardiomyocytes purity. Immunofluorescence and DAPI staining were used to distinguish between cardiomyocytes and other cells. CTnI (green) and nucleus (blue) were stained in cardiomyocytes, and only the nuclei (blue) were stained in other cells. (magnification, 100×)

Fig. 2. Lentiviral transfection of in vitro cardiomyocytes. a MOI = 1, b MOI = 10, c MOI = 50. Transfection efficiency: a: 3.21 ± 0.19%, b: 16.72 ± 2.03%, c: 89.32 ± 8.35%

Fig. 3. H2O2-induced in vitro cardiomyocyte apoptosis. Cardiomyocytes were treated with 100 μmol/L H2O2 (a), 200 μmol/L H2O2 (b), 400 μmol/L H2O2 (c). Data were expressed as mean ± standard deviation

Fig. 4. Flow cytometry of apoptosis. Compared with NC group, a P < 0.05. H2O2 + miR-486 up group compared with H2O2 + UC group, b P < 0.05. miR-486 down group was compared with DC group, c P < 0.05. NC group: negative control group, H2O2 + UC group: H2O2 + miR-486 up control group, DC group: miR-486 down control group

Fig. 5. TUNEL staining. Red fluorescence referred to TUNEL-positive cells, DAPI (blue) referred to nuclear counterstaining (magnification 100×). ap < 0.05 compared with NC group. bp < 0.05: compared with H2O2 + UC group, cardiomyocyte apoptosis index was lower in the H2O2 + miR-486 up group. cp < 0.05: compared with DC group, cardiomyocyte apoptosis index was increased in the miR-486 down group

Fig. 6. Fluorescence-based quantitative PCR results of miR-486 expression. Compared with NC group, a P < 0.05. H2O2 + miR-486 up group compared with H2O2 + UC group, bP < 0.05. miR-486 down group compared with DC group, c P < 0.05

Fig. 7. Fluorescence-based quantitative PCR results of p53 mRNA expression. Compared with NC group, a P < 0.05. H2O2 + miR-486 up group compared with H2O2 + UC group, bP < 0.05. miR-486 down group compared with DC group, c P < 0.05

Fig. 8. Fluorescence-based quantitative PCR results of Bbc3 mRNA expression. Compared with NC group, a P < 0.05. H2O2 + miR-486 up group was compared with H2O2 + UC group, bP < 0.05. miR-486 down group compared with DC group, c P < 0.05

Fig. 9. Fluorescence-based quantitative PCR results of BCL-2 mRNA expression. Compared with NC group, a P < 0.05. H2O2 + miR-486 up group compared with H2O2 + UC group, bP < 0.05. miR-486 down group compared with DC group, c P < 0.05

Fig. 10. Western blot results of relative expression of p53 protein. Compared with NC group, a P < 0.05. H2O2 + miR-486 up group compared with H2O2 + UC group,bP < 0.05. miR-486 down group compared with DC group,c P < 0.05

Fig. 11. Western blot results of relative expression of Bbc3 protein. Compared with NC group, a P < 0.05. H2O2 + miR-486 up group compared with H2O2 + UC group,bP < 0.05. miR-486 down group compared with DC group,c P < 0.05

Fig. 12. Western blot results of relative expression of BCL-2 protein. Compared with NC group, a P < 0.05. H2O2 + miR-486 up group compared with H2O2 + UC group,bP < 0.05. miR-486 down group compared with DC group,c P < 0.05