2 7 dichlorofluorescein diacetate dcfh da

Medium 199 (M-199), fetal bovine serum (FBS), and an antibiotic solution were purchased from Gibco (Grand Island, NY, USA). 2,2-Diphenyl-1-1-picrylhydrazyl (DPPH), rutin, and luteolin were purchased from Sigma-Aldrich (St. Louis, MO, USA). 2′, 7′-Dichlorofluorescein diacetate (DCFH-DA) and radioimmunoprecipitation assay (RIPA) lysis buffer were purchased from Solarbio (Beijing, China). Other materials used are specified in detail in the following sections.

2,7-dichlorofluorescein diacetate (DCFH-DA) (Solarbio, Beijing, China) is a cell-permeable, peroxide-sensitive fluorescent probe that was used to detect intracellular free radicals. Once DCFH-DA entered the cells and hydrolyzed to DCFH in the presence of free radicals such as H₂O₂ and peroxides, DCFH was oxidized to the green, fluorescent product dichlorofluorescein (DCF) and trapped in the cell compartment, where it could be measured by a fluorescence spectrophotometer. The wavelength of the laser was 488–525 nm (excitation and emission, respectively).
In the present study, two GC groups were collected and incubated with DCFH-DA at 37 °C for 20 min. Then, fluorescent DCF was measured using a fluorescence spectrophotometer, and the relative absorbance values that characterize the reactive oxygen species (ROS) content were measured.

TMZ and L-GSH were purchased from Shanghai Macklin Biochemical Technology Co., Ltd. (Shanghai, China). Methylene blue (MB) and 2,7-dichlorofluorescein diacetate (DCFH-DA) were obtained from Beijing Solarbio Science & Technology Co., Ltd. (Beijing, China). Dopamine hydrochloride (DA‧HCl), 5,5-dithiobis (2-nitrobenzoic acid) (DTNB), and indocyanine green (ICG) were purchased from Aladdin (Shanghai, China). 4’,6-Diamidino-2-phenylindole (DAPI), Lyso-Tracker Red, and Hoechst 33,342 were obtained from Beyotime (Shanghai, China). D-luciferin potassium salt was purchased from BioFroxx (Germany). Calcein-AM/PI double staining kit was purchased from Yeasen Biotech Co., Ltd. (Shanghai, China). Annexin V-FITC/PI detection kit was purchased from Becton, Dickinson and Company (USA). The Luciferin (Luc) lentiviral vector was purchased from Shanghai Jikai Gene Chemical Technology Co., Ltd (GV260 vector, Shanghai, China). MGMT enzyme linked immunosorbent assay (Elisa) kit was obtained from Jiangsu Meimian Industrial Co., Ltd (Jiangsu, China).