Figure 4.

Treatment of ASCs with Tat or Nef resulted in higher oxidative stress and mitochondrial dysfunction. (A) After 15 days of treatment with Tat or Nef in ASCs, isolated from different abdominal SCAT healthy donors, reactive oxygen species (ROS) production was assessed spectroscopically by measuring the nitro blue tetrazolium (NBT) absorbance (normalized against protein content) and the CM-H₂DCFDA fluorescence (normalized against DAPI) (n = 9, in duplicate). (B) Mitochondrial mass was evaluated using MitoTracker Red dye. The cationic dye JC-1 was used to evaluate the mitochondrial membrane potential. The fluorescence results are expressed as the mean ± SEM % JC-1 aggregate/monomer ratio, relative to control cells (n = 9, in duplicate). * P < 0.05, ** P < 0.01, *** P < 0.001 vs. control cells.