Figure 5.

Metformin increases dysfunctional mitochondria clearance and inhibits ROS degeneration. To activate AMP-activated protein kinase (AMPK), SHSY5Y cells were preincubated with MET (2mM) for 1 hour, and MPP⁺ (200 μM) was added for 48 hours. To block AMPK or autophagy, SHSY5Y cells were preincubated with Compound C (CC) (10 μM) for 30 minutes or with 3-Methyladenine (3MA) (5mM) for 3 hours before MET addition. (a) After drug treatment, cells were stained with Mitotracker green and Mitotracker deep red for 30 minutes and analyzed by flow cytometry. Photos of ROS fluorescence (b) and intensity of intracellular ROS fluorescence (c). Blue color represents nuclear and green color represents ROS. Scale bar, um. Data are presented as the mean ± SEM of 3 independent experiments. Scale bar, 10 μm. ^* P<.05, ^** P<.01, and ^*** P<.001 vs untreated cells. ^# P<.05, ^## P<.01, and ^### P<.001 vs MPP⁺-treated cells. ^&& P<.01 and ^&&&P<.001 vs MPP⁺/MET co-treated cells. MET, metformin.