Phenylmethanesulfonyl fluoride pmsf

Roxadustat (FG-4592) was purchased from Selleck (Shanghai, China) and dissolved in a mixed solvent containing 5% DMSO, 40% PEG-300, 5% Tween-80 and 50% ddH₂O according to Selleck in vivo usage instructions for mouse intraperitoneal injection. Rox powder was stored at −20 °C in silver paper to avoid light and ensure dryness, with its solvent prepared immediately prior to use. Rox was dissolved in solvent to reach a concentration of 100 mM and then diluted to concentrations of 5–100 μmol/L with cell culture medium for cell assays. METH was obtained from Shanghai Standard Biotech Co., Ltd. (Shanghai, China) and diluted with saline for i.p. injection (2.0 mg/kg). Morphine was purchased from Shenyang First Pharmaceutical Factory (Shenyang, China) and dissolved in saline for i.p. injection (5.0 or 20.0 mg/kg). Deferiprone (DFP) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Phenylmethanesulfonyl fluoride (PMSF) was purchased from Cell Signaling Technology (Danvers, MA, USA). Other common reagents were purchased from Sinopharm (Beijing, China).

Ssb1 (Desit, Chengdu, China), PLGA (lactide/glycolide = 50/50; MW: 94 000 Da; MedChemExpress, USA), polyvinyl alcohol (PVA; BBI Co., Ltd., Shanghai, China), 2-(N-morpholine)ethanesulfonic acid (MES; Heowns Biochem LLC, Tianjian, China), KMnO₄ (Sinopharm Chemical Reagent Co., Ltd., Shanghai, China), radio-immunoprecipitation assay (RIPA) buffer (Solarbio, Beijing, China), phenylmethanesulfonyl fluoride (PMSF, 1:100, Cell Signaling Technology, MA, USA), and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT; Solarbio, Beijing, China) were purchased and used as received. TUNEL, DCFH-DA and DiR iodide were purchased from Beyotime Ltd. (Shanghai, China). Fetal bovine serum (FBS), trypsin-EDTA solution (0.25 %), and Dulbecco’s modified Eagle medium (DMEM) were obtained from Gibco (Burlington, Canada). All other chemicals and reagents of the highest purity available were obtained from commercial sources.