Human Colon Cancer Cell Culture

Human colon cancer HCT116 (kindly provided by Prof. Ber Vogelstein, Johns Hopkins University, Baltimore, MD, USA) and RKO cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) (Life Technologies-Invitrogen, Eggenstein, Germany), supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Corning Life Sciences, New York, NY, USA), plus glutamine and antibiotics (Corning Life Sciences, New York, NY, USA) in a humidified atmosphere with 5% CO₂ at 37 °C. Cells underwent routine testing to ensure that they were mycoplasm negative. The inhibitor of the antioxidant response Brusatol (Sigma-Aldrich, St Louis, MO, USA) [34 (link),35 (link)] was used at 100 µM for 4 h pre-treatment, as previously reported [36 (link)]; Bip/GRP78 inhibitor HA15 (Sigma-Aldrich, St Louis, MO, USA, SML2118) [37 (link),38 (link)] was used at 10 µM for 1 h pre-treatment, as previously reported [20 (link)].