Mitosox red staining kit

Primary antibodies of neuron specific enolase (NSE), light chain 3 (LC3)b, Beclin-1, p62, ATG5 and β-actin were purchased from Abcam (Cambridge, MA, USA); 2′,7′-dichlorofluorescin diacetate (DCFDA) Staining kit, western blotting, secondary antibody labeled with Alexa Fluor-555, 4′,6-diamidino-2-phenylindole (DAPI) and protein extraction kits were bought from Beyotime Institute of Biotechnology (Haimen, China); Cell culture reagents, including Neurobasal-A, fetal bovine serum (FBS) and 0.25% trypsin, were obtained from Thermo; Cell Counting Kit-8 (CCK-8) was purchased from Dojindo Molecular Technologies, Inc. (Kumamoto, Japan); MitoSOX Red Staining Kit was purchased from Invitrogen (Thermo Fisher Scientific, Inc., Waltham, MA, USA).

Mitochondrial ROS production was analyzed using a MitoSOX Red staining Kit (Invitrogen, Carlsbad, CA). The MMP was detected using a MitoTracker Red CMXRos Kit (Invitrogen). Representative images were captured using a TCS SP8 CARS confocal fluorescence microscope (Leica Microsystems). These experiments were repeated three times.