Fig. 1.

MnTE-2-PyP protected from radiation and hyperglycemia-induced mitochondrial damage in normal prostate fibroblasts. Human prostate fibroblast cells were treated with 20 mM glucose (HG) and 30 μM MnTE-2-PyP (T2E) followed by 3 Gy of X-rays (RAD). 5 days after radiation, cells were harvested. A. Mitochondrial and whole cell proteins were extracted and immunoblotted for mitochondrial NRF2, total PGC1α and TOMM20. Quantification of 3 independent western blots is shown below. B. Mitochondrial localization of NRF2. Left: Representative images (20X) of human prostate fibroblasts stained with Mitotracker Red (red) and NRF2 (green). Scale bar = 100 μm. Right: Upper two images represent Mitotracker Red (red) and NRF2 (green) in 40X magnification. RGT group but not the control group showed co-localization of Mitotracker Red and NRF2 in yellow color (areas indicated by white arrows). Scale bar = 50 μm. Right lower panel represents a confocal image (63X) of colocalization between (white signals, analyzed by Image J) TOMM20 (green) and NRF2 (red) in the RGT group. Scale bar = 25 μm. C. Representative images of mitochondrial ATP synthase protein expression. Mean fluorescent intensity of ATP synthase staining per cell is shown below. Scale bar = 100 μm. All figures are representative of at least 3 independent experiments. CON = control, HG = high glucose, T2E = 30 μM MnTE-2-PyP treated, TG = T2E + HG treated, RAD = 3 Gy X-ray treated, RG = RAD + HG treated, RT = RAD + T2E treated and RGT = RAD + HG + T2E treated.(*), ($), (#) and (@) denote a significant difference (p ≤ 0.5) as compared to control, HG, RAD and RAD + HG group respectively. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)