Crm1 binds to and exports Mps1 from the nucleus after mitosis. (A) Export of Mps1 from nucleus upon mitosis completion. SW480 cells were arrested at prometaphase via 100 ng/ml Nocodazole treatment and released into fresh medium for the duration indicated, before being fixed for immunofluorescence staining with anti-Mps1 N1 antibody. DNA was counterstained with DAPI. (B) Mps1 relocated into the nucleus upon LMB treatment. Asynchronized SW480 cells were treated with 10 μM LMB and fixed for immunofluorescence staining at the indicated time points. (C)YFP-fused Mps1 show same pattern in SW480 upon LMB treatment. Asynchronized SW480 cells expressing YFP-Mps1 were treated with 10 μM LMB and fixed for immunofluorescence staining at the indicated time points. (D) Interaction of Mps1 and Crm1. 293 T cells were transiently co-transfected with pEXL-FTH-Crm1 and pRK5-myc-Mps1 and collected for the immuno-precipitation assay. Nonspecific binding product is indicated as symbol *. (E, F) Reciprocal association of Mps1 and Crm1: The bead bound GST-Mps1 was co-incubated with His-crm1 for 3 hrs in lysis buffer and then was collected by centrifuge. The bead bound protein and the proteins in supernatant were examined by Western-blot with antibodies indicated. Reciprocally, bead bound His-Crm1 was incubated with a non-tagged Mps1. The interaction was also determined in the same standard protocol. DAPI, 4′,6-diamidino-2-phenylindole.