Escherichia coli bl21 cells

Dimethyl sulfoxide (DMSO), 1,4-NQ (98% purity determined by gas chromatography) and anti-GAPDH antibody were from Wako Pure Chemical Industries (Osaka, Japan), Tokyo Chemical Industries (Tokyo, Japan) and Santa Cruz Biotechnology (Santa Cruz, CA, USA), respectively. Biotin-PEAC₅-maleimide (BPM) and Na₂S₄ were from Dojindo Laboratories (Kumamoto, Japan). Dynabeads M-280-sheep anti-rabbit immunoglobulin G (IgG) was from Invitrogen (Carlsbad, CA, USA). Anti-Akt, anti-CREB, anti-phosphorylated Akt (Ser473), anti-phosphorylated CREB (Ser133), horseradish peroxidase (HRP)-conjugated anti-biotin antibodies, anti-rabbit antibodies and anti-mouse IgG secondary antibodies were from Cell Signaling Technology (Beverly, MA, USA). Escherichia coli BL21 cells and trypsin were from Promega Co. (Madison, WI, USA). Glutathione 4B Sepharose was from GE Healthcare (Chicago, IL, USA). All other reagents were of the highest purity available.

MeHg and avidin-agarose was obtained from Sigma-Aldrich (St. Louis, MO, USA). Escherichia coli BL21 cells and trypsin were purchased from Promega Co. (Madison, WI, USA). Anti-GAPDH antibody and anti-Lamin B1 antibody were obtained from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA). Anti-G6PD antibody, BPM, 4,6-diamidino-2-phenylindole (DAPI), annexin V fluorescein isothiocyanate (FITC), and propidium iodide were obtained from Bethyl Laboratories (Montgomery, TX, USA), Dojindo (Kumamoto, Japan), Nacalai Tesque Inc. (Kyoto, Japan), MBL (Aichi, Japan), Wako Pure Chemical Industries (Osaka, Japan), respectively. Anti-Akt, anti-CREB, anti-phosphorylated Akt (Ser473), anti-phosphorylated CREB (Ser133), anti-Bcl-2, horseradish peroxidase (HRP)-conjugated anti-rabbit antibodies, LY294002, and wortmannin were purchased from Cell Signaling Technology (Beverly, MA, USA). Alexa 488-secondary antibody, rhodamine phalloidin and Lipofectamine 2000 were obtained from Thermo Fisher Scientific (MA, USA). All other reagents used were of the highest purity available.