Intracellular ros detection kit

The intracellular ROS levels of cells were measured using an intracellular ROS detection kit (Sigma-Aldrich) according to the manufacturer’s instructions. Briefly, the cells were inoculated into a 96-well microtitration plate, 100 µL of ROS detection reagent mixture was added, and the plate was incubated for 1 h at 37 °C in a 5% CO₂ humidified incubator. Subsequently, the data were collected using a fluorescence microscope and a multifunctional microplate reader.

Desktop high-speed refrigerated centrifuge (Allegra 64R High Speed Centrifuge, Beckman, USA); flow cytometry (Navios; Beckman, USA); multi-function microplate reader (Synergy; Biotech, USA); inverted fluorescence microscope (DM1LLED; Leica, Germany); laser confocal microscopy (LSM710; ZEISS, Germany); DMEM medium, fetal bovine serum, and trypsin (Gibco, New Zealand); H₂O₂ (Chengdu Jinshan Chemical Reagent Co., Ltd., China); CCK-8 solution (Japan Tongren Chemical Research Institute, China); annexin V-FITC/PI double-stained cell apoptosis detection kit (BD; San Diego, USA); intracellular ROS detection kit (Sigma, USA); JC-1 cell mitochondrial membrane potential detection kit and TUNEL apoptosis detection kit (Nanjing Kaiji Biotechnology Co., Ltd., China); DAPI and BCA protein assay kit (Beijing Suobao Biotechnology Co., Ltd., China); superoxide disproportionation enzyme detection kit and catalase detection kit (Nanjing, China); and malondialdehyde detection kit (Shanghai Biyuntian Biotechnology Co., Ltd., China).