Figure 1

Nup88 and Nup214 form a complex during mitosis. (A) Schematic diagrams of the structure of the nuclear pore complex (upper) and the nucleoporin Nup88 domains organization (lower). (B) Immunoprecipitates from mitotic HeLa cell extracts with anti-Nup214, anti-Nup88 antibodies or nonspecific rabbit antibodies (IgG) were analyzed by SDS/PAGE, followed by immunoblotting with an anti-Nup88, anti-Nup214 or anti-Nup153 antibodies respectively. In the lanes marked ''input'', 20 μl of the 500-μl extract that was used per immunoprecipitation was analyzed directly. (C) Immunoprecipitates from mitotic HeLa cell extracts with anti-Nup88 antibodies or nonspecific rabbit antibodies (IgG) were analyzed by SDS/PAGE, followed by immunoblotting with an anti-CENP-E antibody (upper panel). HeLa cells were costained with anti-Nup88 (green) and anti-CENP-E (red) antibodies and analyzed by confocal laser microscopy. Chromatin was visualized using DAPI (blue)(Lower panel). (D) HeLa cells were costained with anti-Nup88 (green) and anti-Nup214 (red) antibodies and analyzed by confocal laser microscopy. Chromatin was visualized using DAPI (blue). (E) HeLa cells were costained with anti-α-tubulin (green) and anti-Nup88 (red) antibodies and analyzed by confocal laser microscopy. Chromatin was visualized using DAPI (blue). Scale bar, 10 μm.