FCCP and Valinomycin reduce HSC ATP levels, ROS production and stimulate mitochondrial biogenesis related genes. (A) Cells were treated with 5 μM of FCCP or Valinomycin for 24 hours and ATP levels were measured using a luciferase based ATP determination kit. (B) ROS levels were analyzed in mHSCs treated for different time points with chemical uncouplers by DCF fluorescence. HSCs were treated with 5 μM of FCCP or Valinomycin for 7 days and the expression of genes involved in mitochondrial biogenesis PGC-1α (C), Tfam, Ndufs7 and CoxIV (D) were analyzed by RT-qPCR. * indicates P < 0.05 compared to control groups. Data are expressed as means of 3 independent experiments ± SEM.