FIG. 3.

Sensitivity of IRP1 and IRP2 cysteine mutants to diamide. Flp-In TREx HEK293 cells stably expressing FLAG-tagged IRPs were treated with tetracycline overnight to induce protein expression. (A) Cell lysates (10 μg) were treated with increasing concentrations of diamide, with or without β-ME (IRP1, 1%; IRP2, 0.5%) prior to EMSA. Quantification of three independent experiments is shown. Error bars represent the standard errors of the means. (B) Cell lysates (10 μg) were untreated (−) or treated with 0.5 mM diamide (D), 0.5 mM diamide followed by β-ME (DM), or β-ME alone (M) prior to EMSA. Only FLAG-supershifted IRP-IRE complexes are shown.