FIGURE 5.

Effects of miR-27 on mitochondrial content, membrane potential, and morphology. ASC were transduced with Lenti/miR-Control (miR-Cont), Lenti/miR-27a (miR-27a), or Lenti/miR-27b (miR-27b). Three days later, the cells were induced to adipocyte differentiation with adipogenic inducers for a period from days 0 to 14. A, the relative mtDNA content was evaluated by a ratio of the DNA level of mitochondrial ND1 to that of nuclear 18 S rRNA. The relative mtDNA content in ASC overexpressing Lenti/miR-Control at day 0 was set to 1. B and C, the mitochondrial membrane potential of ASC was measured by detecting the accumulation of TMRE, a red fluorophore, in active mitochondria by flow cytometry. The cells without the addition of TMRE were used to facilitate the gating of red events. A statistical analysis of the percentage of TMRE-positive cells was shown in C. D, ASC at days 0 and 14 were stained with MitoTracker Red. The mitochondrial morphology was analyzed a confocal microscopy system. Scale bar = 10 μm. The bar graph represents the percentage of cell populations with fragmented mitochondria. *, p < 0.05; **, p < 0.01 compared with Lenti/miR-Control at day 0. *, p < 0.05; ##, p < 0.01 compared with Lenti/miR-Control at day 14.