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Cell lines and culture conditions HepG2 cells, a human hepatoma cell line, were obtained from the American Type Culture Collection (Manassas, VA) and were cultured in Dulbeccos modified Eagles medium (DMEM) containing 10% (v/v) fetal bovine serum, 2 mM glutamine, 5 U/mL penicillin, and 50 g/mL streptomycin at 37 C in a humidified O 2 /CO 2 (19:1) atmosphere

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The GSH-containing syringe of the stopped-flow spectrofluorometer was supplemented again with 0.4 U/mL GR and 0.25 mM NADPH to remove trace amounts of GSSG and to prevent the thermodynamically favored reverse reaction 32

Furthermore, heme can drive TNF-- and ROS-mediated necroptosis [225], heme oxygenase 1-dependent ferroptosis [226], and NLRP3 inflammasome-associated pyroptosis [227], which are highly proinflammatory RCDs