Figure 9. Analysis of the effect of the miR-15b – SIRT4 axis on mitochondrial morphology and fragmentation. Human dermal fibroblasts were transfected with control oligos (A) or miR-15b inhibitor oligos in the absence (B) or (C) presence of siRNA duplexes against SIRT4. Four days later, cells were stained with SIRT4 and MTCO2 specific antibodies. Confocal laser scanning microscopy and an ImageJ macro were employed to analyse fold changes in mitochondrial morphology-related parameters, including %area (total mitochondrial area/total cell area), mitochondrial perimeter, circularity, and solidity. Approximately two hundred cells were analysed per experimental condition. To evaluate statistical significance, ANOVA SNK were performed (*p<0.05). Bar: 20 μm.