Mislocalization of the Nup107–160 complex from kinetochores impairs the kinetochore targeting of Crm1 and RanGAP1. (A) Cells were treated for 20 min with 10 ng/ml leptomycin B or for 72 h with control, Seh1, Nuf2 or the ‘combined siRNA' duplexes and subsequently pre-extracted, fixed and probed with anti-RanGAP1 (red) and anti-Crm1 (green) antibodies, CREST serum (blue) and DAPI. Single planes from deconvolved Z-stack images (scale bar, 5 μm) and three-fold enlargements of the boxed areas are presented. (B) Remaining levels of Nup133, RanGAP1 and Crm1 at kinetochores in control cells, upon treatment with Seh1, Nuf2 or the combined RNAi, or after LMB treatment, were quantified based on deconvolved Z-stack images (see Materials and methods).