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To evaluate the effects of compound on GPX4 activity, compound was first pre-incubated with purified GPX4 in the assay buffer [Tris-HCl (100 mM, pH 7.4), EDTA (5 mM), Triton X-100 (0.1% v/v), NADPH (0.2 mM), glutathione (3 mM), and glutathione reductase (1 unit)] for 5 min at 37C