FIG. 3.
Identification of the YB-1-binding domain in IRP2. (A) Schematic illustration of the ThioHis-IRP2 deletion mutants used in this study. (B) Expression of recombinant proteins. Deletion mutants were expressed in bacteria and analyzed by Coomassie brilliant blue staining of SDS-PAGE gels. (C) GST pull-down assay. 1 μg of GST-YB-1 or GST which binds to 30 μl of glutathione-Sepharose 4B beads was incubated with a 100-fold excess of ThioHis-IRP2 deletion mutants. Proteins bound to the beads were resolved by SDS-PAGE and analyzed by Western blotting using an anti-Thio antibody.