Fig. 5

Analysis of oxidative stress determinants during DIPS of HGSOCs. Changes in the production of mitochondrial superoxide (a), cellular peroxides (b), mitochondrial membrane potential (c), and mitochondrial mass (d) quantified according to the fluorescence of MitoSOX Red, DHR, JC-1, and NAO, respectively. As per mitochondrial membrane potential measurements, JC-1 accumulates in the functional mitochondria, forming red fluorescent J-aggregates (at 590 nm), while mitochondria de-energization results in the accumulation of green fluorescent monomers (at 535 nm). The rise in the green/red fluorescence intensity ratio indicates a drop of the mitochondrial membrane potential values. The results are based on 8 independent experiments using HGSOCs obtained from different patients. The results are expressed as the means ± SEM. **p < 0.01; ***p < 0.001 vs. young cells. RFU relative fluorescence units