Figure 4. Association between Nup98-HoxA9 and Crm1 is critical for the Hox Gene activation mediated by Nup98-HoxA9.
(A) Top panel: Nup98-HoxA9 interacts and sequesters Crm1 onto Nup98-HoxA9 dots. HeLa cells were transfected with the EGFP-Nup98-HoxA9 expressing plasmid. After 24 hr, cells were fixed and stained with an anti-Crm1 antibody. Arrows indicate the cells transfected. Bottom panel: Nup98-HoxA9 ES cells were fixed and co-stained with anti-FLAG (M2) and anti-Crm1 antibodies. Merged image of FLAG (green) and Crm1 (red) is shown. Bar, 5 μm. (B) The effect of LMB treatment on the cellular localization of Nup98-HoxA9. Nup98-HoxA9 ES cells were cultured either in the presence or absence of 5 nM LMB for 2 hr, fixed and stained with antibodies against FLAG (M2) and Crm1. Merged images of FLAG (green) and Crm1 (red) are shown. Nuclei were stained with DAPI. Bar, 10 μm. (C) Effect of LMB treatment on the regulation of Hox cluster genes. Nup98-HoxA9 ES cells were cultured in the presence or absence of 5 nM LMB for 3 or 6 hr and the expression of indicated genes was analyzed by qPCR. GAPDH was used as a reference gene. EGFP, enhanced green fluorescent protein; LMB, leptomycin B; qPCR, quantitative polymerase chain reaction.
Figure 4—figure supplement 1. Effect of LMB treatment on the FLAG-Nup98-HoxA9 protein level.
Figure 4—figure supplement 2. Effect of LMB treatment on the cellular localization of various NupFG-HoxA9 fusions.
