Regulation of Connexin 43 Phosphorylation

TNF-α and IL-1β were obtained from Alomone (Jerusalem, Israel); Fasudil, Y-27632, ethidium (Etd⁺) bromide and lanthanum (La³⁺) chloride were obtained from Sigma-Aldrich (St. Louis, MO, USA). TBARS assay kit was obtained from Cayman Chemicals (Ann Arbor, MI, USA), and CellTiter96^® Non-Radioactive Cell Proliferation Assay was obtained from Promega (Madison, WI, USA). The mimetic peptides gap19 (KQIEIKKFK, intracellular loop domain of Cx43) and TAT-L2 (YGRKKRRQRRRDGANVDMHLKQIEIKKFKYGIEEHGK, second intracellular loop domain of Cx43) were obtained from GenScript (Piscataway Township, NJ, USA) [25 (link)]. The monoclonal anti-α-tubulin antibody was purchased from Sigma-Aldrich (St. Louis, MO, USA). The polyclonal anti-phosphorylated-MYPT1 (Thr696) antibody was obtained from Merck Millipore (Darmstadt, Germany). The monoclonal anti-MYPT1 antibody was purchased from BD Transduction Laboratories (San José, CA, USA). The monoclonal anti-unphosphorylated Cx43 antibody was obtained from Invitrogen (Carlsbad, CA, USA). Anti-mouse and anti-rabbit secondary antibodies conjugated to horseradish peroxidase were from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA).