Cell Cycle Analysis with Ros Scavengers

The cell cycle was analyzed as described previously [62 (link)]. Ros scavengers used were: α-tocopherol (100 µM, Vit. E) or Ferrostatin-1 (1 µM). The commercial inhibitors used were: Z-IETD-FMZ (50 µM: Selleckchem, Houston, TX, USA), Z-LEHD-FMK (50 µM: Selleckchem, Houston, TX, USA) and Oxythiamine chloride hydrochloride (OT, 1 and 10 mM: Santa Cruz Biotechnology, Heidelberg, Germany). The cells were seeded in culture plates and pre-treated with commercial inhibitors for 1.5 h before they were exposed to HCA for different times. Caspase-3/7 activity was measured using the Caspase-Glo^® 3/7 assay according to the manufacturer’s instructions (Promega, Madison, WI, USA).