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Membranes were blocked with 5% non-fat milk in Tris-buffered saline with 0.1% Tween-20 (TBST) for 1 h at room temperature and then incubated overnight at 4C with primary antibodies specific for GPX4 (1:1,000, ab125066, Abcam, Britain), NCOA4 (1:1,000, ab86707, Abcam, Britain), FTH-1 (1:1,000, ab75973, Abcam, Britain), TFR-1 (1:1,000, ab214039, Abcam, Britain), SLC7A11 (1:1,000, ab175186, Abcam, Britain), and -actin (1:10,000, AF7018, Affinity, United States)

USP33 K D = 7 M