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IRP2 is subject to iron-dependent oxidation in vitro. Purified IRP1 or IRP2 was incubated with 10 mM DTT and FeCl3 at the indicated concentrations for 4 h at 37°C. Oxidative modifications were detected by the carbonyl assay (A), and total protein was detected by Coomassie staining (B). A time course for oxidation of IRP2 shows carbonyl modifications acquired during treatment with 5 μM FeCl3 and 10 mM DTT (C).
