Figure 3. Effect of simvastatin on reactive oxygen species (ROS) levels and nuclear factor (erythroid-derived 2)-like 2 (NRF2) translocation. (A) Flow cytometric distribution of cells according to their cellular (DCF) and mitochondrial matrix (Mitosox) ROS content in a reference experiment to define gates relative to low and high ROS respectively. (B) Percentage of MitoSOX positive human dermal fibroblasts with elevated mitochondrial superoxide anion (●) and DCF negative cells with low cellular reactive oxygen species (○) after exposure to different concentrations of simvastatin for 72 h. Data (n=9) are reported as mean % of cells and standard error. (C) Nuclear translocation of NRF2 in simvastatin treated samples for 72 h (20 and 40 nM) and unexposed control. Data are expressed as ratio of NRF2-FITC associated green fluorescence in the nucleus and cytoplasm. Data are reported as box-plot with 50% of the population reported in the box; horizontal lines indicate min, median, and max values. Significance difference vs 0 nM + p<0.05, ++ p<0.01, +++ p<0.001, ++++ p<0.0001.