Fig. 4.

Increase of mitoROS but not reduction of cytosolic ROS involves in MGO-induced cell death in ARPE-19 cells.

(A) Cells were 30 min pretreated with NAC (3 and 10 mM) prior to MGO (300 μg/ml) treatment for 6 h. Then Cell viability was determined. (B, C, E, F) Cells were 30 min pretreated with metformin (6 mM) and/or NAC (3 mM) followed by MGO (300 μg/ml) treatment for 4 h. DCFDA (B), DHE (C), mitoSOX (E), and mitoPY1 (F) were used to measure ROS. (D) Cells were 30 min pretreated with DPI (3 μM) and/or metformin (6 mM) prior to MGO (300 μg/ml) treatment for 6 h. Cell viability was measured. (G) Cells were treated with NAC (5 mM) and immediately subjected into Seahorse XFe24 analyzer for measurement of OCR as described in Fig. 3D. Data were the mean ± S.E.M. of at least 3 independent experiments. *p < 0.05, indicating the significant effect of MGO; #p < 0.05, indicating the reversal effects of metformin and NAC. **p < 0.05, indicating the significant reduction of ROS by NAC.