ITCs (SFN, 6-MSITC, and 6-MTITC) were purified from Wasabi by reversed-phase high performance liquid chromatography (HPLC) to >99.3% purity26 (link) and dissolved in dimethyl sulfoxide for cell culture experiments. The antibodies against Nrf2 (C-20), Keap1 (E-20), NQO1 (C-19), HSP70 (D69), GAPDH, rabbit IgG, and horseradish peroxidase (HRP)-conjugated anti-goat secondary antibody were purchased from Santa Cruz Biotechnology. AKR1C1, AKR1C3, and TXNRD1 antibodies were obtained from Abcam. HRP-conjugated anti-rabbit and anti-mouse secondary antibodies were from Cell Signaling Technology.
IMR-32 cell culture. Human neuroblastoma IMR-32 cells (cell no. TKG0207) were obtained from Riken Bioresource Center Cell Bank. IMR-32 cells were grown in Eagle’s Minimum Essential Medium (Nissui Seiyaku) supplemented with 2 mM l-glutamine (Nacalai Tesque), 1% v/v MEM nonessential amino acid solution (Nacalai Tesque), and 10% v/v fetal bovine serum (Equitech-Bio) under a humidified 5% CO2 atmosphere at 37 °C.
IMR-32 cell culture. Human neuroblastoma IMR-32 cells (cell no. TKG0207) were obtained from Riken Bioresource Center Cell Bank. IMR-32 cells were grown in Eagle’s Minimum Essential Medium (Nissui Seiyaku) supplemented with 2 mM l-glutamine (Nacalai Tesque), 1% v/v MEM nonessential amino acid solution (Nacalai Tesque), and 10% v/v fetal bovine serum (Equitech-Bio) under a humidified 5% CO2 atmosphere at 37 °C.