Figure 2
From: miR-27 regulates mitochondrial networks by directly targeting the mitochondrial fission factor
miR-27 regulates the mitochondrial fission factor (MFF) expression. After transfection of pre-miR-27, anti-miR-27 and control miRNA, the MFF mRNA and protein levels were analyzed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) (a) and western blotting (b), respectively. (a) The relative expression of MFF mRNA was analyzed using GAPDH mRNA for normalization. The data represent the mean±s.e.m. from three independent experiments. (b) The MFF protein level was analyzed by western blotting using an anti-MFF antibody, and the β-actin level is shown as a loading control. Images are representative of three independent experiments, and the numbers represent the mean±s.e.m. from three independent experiments. (c) Schematic representation of the reporter plasmids pEGFP (control), pEGFP-MFF 3U and pEGFP-MFF 3UM, the last of which bears five mutated nucleotides in the MFF mRNA that correspond to the miR-27 seed region. (d) CHANG liver cells were co-transfected with the plasmids presented in panel c and with the indicated miRNAs. Forty-eight hours after transfection, the EGFP expression levels were assessed by western blotting. The data represent the mean±s.e.m. of three independent experiments. *P<0.05. EGFP, enhanced green fluorescent protein.
