Fig. 7

miR-484 regulates Fis1-related mitochondrial fission. A The expression of miR-484 in human HCC cell lines (Huh7, HepG2, Hep3B, and SK-Hep1) and normal hepatocyte line (LO2) were detected by RT-qPCR. B Cell proliferation potentials in HepG2 and Huh 7 cells were assessed by CCK8 assay at day 1, 2, 3, 4 and 5 after respectively transfected mimics-NC or miR-484 mimics. C Representative images of colony formation of HCC cells by staining with crystal violet after transfected mimics-NC or miR-484 mimics. Statistical analysis of colony formation assay. D The target genes of miR-484 predicted by miRWalk. E The effect of miR-484 mimics or miR-484 inhibitor on the mRNA expression of Fis1 in HCC cells was detected by qRT-PCR. F The protein expression of Fis1 was assessed in HCC cells after transfection with miR-484 mimics, miR-484 inhibitor or control mimics by Western blotting. G Schematic illustrations showed the alignment of miR-484 and Fis1. The red part was the mutagenesis nucleotides. H The luciferase activity of wild-type and mutant Fis1 was detected by dual Renilla and firely luciferase reporter assays after co-transfected with either the miR-484 or control mimics. I Mitochondrial morphology was evaluated using a confocal microscope after MitoTracker Green staining. The ratio of the morphological changes of mitochondria in per 100 cells in each group was analyzed according to the mitochondrial length. J-L Alterations in pyruvate production, lactate production and ATP production levels were detected in HepG2 and Huh7 cells. (M–N) The ECAR and OCR in HepG2 and Huh7 cells were measured. Data were represented as means ± SEM with at least three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001