Figure 2
From: Antileukemic activity of nuclear export inhibitors that spare normal hematopoietic cells
KPT-185 inhibits CRM1-cargo interactions and nuclear export. (a) GST fusion of the NPMc+(mutA) NES was immobilized on glutathione sepharose and then incubated with 10 μM CRM1 proteins that were preincubated with either buffer or inhibitors (20 μM LMB or 200 μM KPT-185, an analog of KPT-251) and molar excess of RanGTP. After extensive washing, a fraction of the bound proteins was visualized by SDS-PAGE and coomasie blue staining. (b) EGFP-NPM (wild type) is localized in the nucleolus when transfected into HeLa cells, whereas a mutant of NPM found in AML called NPMc+(MutA) is mislocalized to the cytoplasm. Treatment of 10 nM leptomycin B or 100 nM KPT-185 reverted the mislocalization such that EGFP-NPMc+(mutA) is now back in the nucleus.
