Figure 2.

Inhibition of mitochondrial respiration in the presence of RF-EMR is associated with increased ROS production. (A) Spermatogonia-like (GC1) and (B) spermatocyte-like (GC2) cell lines were seeded to glass coverslips overnight and treated with mitochondrial electron transport chain inhibitor rotenone, in the presence or absence of RF-EMR exposure (1.8 GHz, 0.15 W/kg), for periods of up to 6 h. Alongside these experiments, another electron transport inhibitor, antimycin A, was also utilized for GC1 (C) and GC2 cells (D). Mitochondrial ROS production was assessed using the MSR probe. This analysis was again restricted to the live population, determined by co-labeling with SYTOX green vitality stain. Glucose and succinate substrates were utilized for comparison of mitochondrial ROS generation in GC1 (E) and GC2 (F) cells in the presence of RF-EMR. Cells were seeded to coverslips overnight in DMEM media as detailed above, and refreshed with this DMEM media or 5 mM succinate media DMEM (devoid of glucose) for the course of the experiment. These analyses were performed on at least three biological replicates and data are presented as mean ± SEM. **p < 0.01, *p < 0.05 compared to unexposed controls.