ROS amounts were assessed with Cellrox green and Mitosox-red (Thermo Fisher Scientific). Upon treatment and PBS wash, the cells underwent staining with 5 μmol/L Cellrox green or 5 μmol/L Mitosox-red in pre-warmed serum-free DMEM shielded from light for 30 min. Fluorescence intensity was measured using a Leica SP8 confocal fluorescence microscope and a FACS Calibur system. All assays were repeated at least three times.
MMP was examined by assessing Rhodamine 123 (Thermo Fisher Scientific) and TMRM (Thermo Fisher Scientific). Upon treatment, cells underwent incubation with 1 μmol/L Rhodamine 123 or 20 nmol/L TMRM (30 min, 37 °C) in the dark. A Leica SP8 confocal fluorescence microscope and a FACS Calibur system were utilized for analysis. All assays were repeated at least three times.
MMP was examined by assessing Rhodamine 123 (Thermo Fisher Scientific) and TMRM (Thermo Fisher Scientific). Upon treatment, cells underwent incubation with 1 μmol/L Rhodamine 123 or 20 nmol/L TMRM (30 min, 37 °C) in the dark. A Leica SP8 confocal fluorescence microscope and a FACS Calibur system were utilized for analysis. All assays were repeated at least three times.