Ethidium homodimer 2 ethd 2

Agarose was acquired from OmniPur (Merck, Darmstadt, Germany). SYLGARD 184 silicone elastomer kit, including poly(dimethylsiloxane) (PDMS) base and catalyst, was purchased from Dow Corning (Auburn, MI). Di-2-pyridylketone-4,4-dimethyl-3- thiosemicarbazone (Dp44mT), ribonuclease A (RNase A), and propidium iodide (PI) were purchased from Sigma-Aldrich (St. Louis, MO). Dulbecco’s modified Eagle’s medium (DMEM), Dulbecoco’s phosphate-buffered saline (DPBS), fetal bovine serum (FBS), and penicillin-streptomycin mixtures (antibiotics) were from Gibco BRL (Carlsbad, CA). Glioma cell line U251 was from ATCC (Manassas, VA). SYTO 10 and ethidium homodimer-2 (EthD-2) were purchased from Invitrogen (Waltham, MA).

Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS) were obtained from Gibco (Grand Island, NY, USA). Bilirubin, ethidium bromide (EtBr), Hoechst 33342, 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT), human serum albumin (HSA), and trypan blue were purchased from Sigma-Aldrich, Co. (St. Louis, MO, USA). Ethidium homodimer-2 (EthD2) was purchased from Invitrogen (Thermo Fisher Scientific). VX-765 was purchased from Selleck Chemicals (Houston, TX, USA). Anti-caspase-1 antibody (ab1872) and anti-NLR family pyrin domain containing 3 (NLRP3) (ab91413) were purchased from Abcam (Cambridge, UK). Anti-glial fibrillary acidic protein (GFAP) antibody (3670), anti-β-actin (3700), an HRP-conjugated anti-rabbit secondary antibody (7074P2), and an HRP-conjugated anti-mouse secondary antibody (7076P2) were purchased from Cell Signaling Technology. IL-1β enzyme-linked immunosorbent assay (ELISA) kit and IL-18 ELISA kit were from USCN Life Science, Inc. (Wuhan, China). Lactate dehydrogenase (LDH) cytotoxicity detection kit was obtained from Beyotime Biotechnology (Shanghai, China). All other chemicals and reagents were from Beyotime Biotechnology unless otherwise specified.