Ethidium etd bromide

HEPES, water (W3500), Dulbecco′s Modified Eagle Medium (DMEM), 4,6,-O-ethylidene-D-glucose (ETDG), anti-GFAP polyclonal antibody, cytosine arabinoside (Ara-C), LaCl₃ (La³⁺), cytochalasin B (Cyto-B) and ethidium (Etd) bromide were purchased from Sigma-Aldrich (St. Louis, MO, USA). Fetal bovine serum (FBS) was purchased from Hyclone (Logan, UT, USA), whereas penicillin, 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose (2-NBDG), streptomycin, Propidium (Prd) iodide, diamidino-2-phenylindole (DAPI), YO-PRO-1, goat anti-mouse Alexa Fluor 488/555 and goat anti-rabbit Alexa Fluor 488/555 were from Thermo Fisher Scientific (Waltham, MA, USA). IL-1β and TNF-α were obtained from Roche Diagnostics (Indianapolis, MI, USA). Normal goat serum (NGS) was purchased from Zymed (San Francisco, CA, USA). Anti-Cx43 monoclonal antibody (610061) was obtained from BD Biosciences (Franklin Lakes, NJ, USA). The mimetic peptides gap19 (KQIEIKKFK, intracellular loop domain of Cx43), gap19^I130A (KQAEIKKFK, negative control), Tat-L2 (YGRKKRRQRRR-DGANVDMHLKQIEIKKFKYGIEEHGK, intracellular loop domain of Cx43), and Tat-L2^H126K/I130N (YGRKKRRQRRR-DGANVDMKLKQNEIKKFKYGIEEHGK, negative control) were obtained from Genscript (Piscataway, NJ, USA).

Ethidium (Etd) bromide, LaCl₃, adenosine triphosphate disodium (Na₂ATP), cyclopiazonic acid (CPA, sarcoplasmic reticulum Ca²⁺-pump inhibitor), carbonyl cyanide 3-chlorophenylhydrazone (CCCP, H⁺ ionophore and uncoupler of oxidative phosphorylation in mitochondria), U73122 [Phospholipase C (PLC) inhibitor], carbenoxolone (CBX) and streptomycin were obtained from Sigma-Aldrich (St. Louis, MO, USA). Fura-2-AM was obtained from Molecular Probes (Eugene, OR, USA) and gentamicin sulfate from Invitrogen Life Technologies (Carlsbad, CA, USA). Polyclonal anti-P2Y₂, -P2Y₄, and -P2Y₆ receptor antibodies, as well as goat anti-rabbit and anti-mouse secondary antibodies conjugated to horseradish peroxidase, were obtained from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA). The mimetic peptide Gap26 (sequence: N-VCYDKSFPISHVR-C) was synthesized by Beijing SBS Genetech Co. Ltd. (Beijing, China).

The mimetic peptides gap19 (KQIEIKKFK, intracellular loop domain of Cx43), TAT-L2 (YGRKKRRQRRRDGANVDMHLKQIEIKKFKYGIEEHGK, second intracellular loop domain of Cx43) and ¹⁰panx1 (WRQAAFVDSY, first extracellular loop domain of Panx1) were obtained from GenScript (Piscataway Township, NJ, USA). Ethanol was obtained from Merck Millipore (Darmstadt, Germany). HEPES, ns-398, sc-560, L-N6, SB203580, lanthanum (La³⁺), anti-glial fibrillary acidic protein (GFAP) monoclonal antibody, BAPTA-AM, carbenoxolone (CBX), ethidium (Etd) bromide and probenecid were purchased from Sigma-Aldrich (St. Louis, MO, USA). Goat anti-mouse Alexa Fluor 488/555 and Hoechst 33342 were obtained from Thermo Fisher (Waltham, MA, USA). Normal goat serum (NGS) was purchased from Zymed (San Francisco, CA, USA).