Fig. 4

CV-exo-miR-2904 inhibits hepatocytes proliferation and promotes apoptosis in vitro. A Expression of CV-exo-miR-2904 in HepG2 cells after overexpression. B CCK-8 assay to determine the effect of CV-exo-miR-2904 on the viability of HepG2 cells. C The relative mRNA expression of proliferation marker genes, cyclinE1, cyclinD1, and cyclinB1, after CV-exo-miR-2904 overexpression in HepG2 cells. D LDH-cytotoxicity assay to detect the effect of CV-exo-miR-2904 overexpression on the cytotoxicity of HepG2 cells. E Flow cytometry results to assess the effect of CV-exo-miR-2904 overexpression on apoptosis. F Quantification of flow cytometry results: Cell apoptosis rate = early cell apoptosis rate (Q2) + late cell apoptosis rate (Q3). G Immunofluorescence staining of apoptosis-related proteins, Caspase3 and P53, in HepG2 cells overexpressing CV-exo-miR-2904. Scale: 1 bar represents 100 μm. H The amount of Caspase3 and P53 was quantified using the ImageJ software. I The relative mRNA expression of apoptosis markers, Caspase3, P21, and P53, after CV-exo-miR-2904 overexpression in HepG2 cells. Data are presented as mean ± SD. ** represents P < 0.05, *** represents P < 0.001, and **** represents P < 0.0001. MN mimic negative control