Hoechst 33 342 live cell staining solution

Discard the medium, wash twice with PBS, add dropwise an appropriate amount of Hoechst 33,342 Live Cell Staining Solution (Beyotime Biotechnology, Shanghai, China), and mix gently. Incubate for 10 min at a temperature suitable for cell culture. Aspirate the staining solution, wash 2–3 times with culture medium or PBS, and observe under the fluorescence microscope.

Glyceryl monostearate, medium chain triglycerides and poloxamer 188 were purchased from AIKE REAGENT Co., Ltd. (Chengdu, China). Absolute ethanol was purchased from the DAMAO Chemical Reagent Factory (Tianjin, China). Phosphoric acid and methanol were supplied by FUYU CHEMICAL (Tianjin, China). TMZ and CUR were purchased from Aladdin Co., Ltd. (Shanghai, China). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was purchased from Med Chem Express Co., Ltd. (New Jersey, USA). Hoechst 33342 Live Cell Staining Solution was obtained from Beyotime Co., Ltd. (Shanghai, China). All other chemical compounds were of analytical purity. Rat glioma cell line (C6) and human brain glial normal cell line (HEB) were obtained from Jennio Biotech Co., Ltd. (Guangzhou, China).

Cell Counting Kit-8 (CCK-8), NADP + /NADPH assay kit, Bicinchoninic acid (BCA) protein assay kit, radioimmunoprecipitation assay (RIPA) lysis buffer, Mito-Tracker Green staining solution and Hoechst 33342 live cell staining solution were purchased from Beyotime (Shanghai, China). ELISA assay kit was purchased from mlbio (Shanghai, China). 4’, 6-Diamidine-2’-phenylindole dihydrochloride (DAPI), coverslips and Triton X-100, penicillin, streptomycin and bovine serum albumin (BSA) was purchased from Solarbio (Beijing, China). Enhanced chemiluminescence (ECL) kits were obtained from Thermo Fisher Scientific Pierce (Rockford, IL, USA). Fetal Bovine Serum (FBS) and Dulbecco’s Modified Eagle’s medium (DMEM) were purchased from Hyclone (Logan, UT, USA). Rotenone was purchased from MCE (Shanghai, China). Mito-SOX red mitochondrial superoxide indicator was purchased from Yeasen (Shanghai, China). Trizol Reagent, cDNA synthesis superMix and Two-step RT-PCR superMix were purchased from TransGen Biotech (Beijing, China). Primary antibodies, including NLRP3, ASC, Caspase-1, IL-1β and α-Tubulin, were purchased from Proteintech (Wuhan, China), and NF-κB p65, Phospho-NF-κB p65, IκBα and Phospho-IκBα were purchased from Cell Signaling Technology (Danvers, MA, USA). Peroxidase-conjugated goat anti-mouse IgG and goat anti-rabbit IgG were purchased from Proteintech (Wuhan, China).