Figure 2

Overexpression of GFP-Nup88 and Nup214 _684-974 rescues spindle bipolarity. (A) HeLa cells were transfected with a GFP-Nup88 expression plasmid. After 48 h, the cells were lysed and analyzed by immunoblotting with an anti-Nup88 antibody. (B) Representative images of mitotic HeLa cells transfected with the plasmid overexpressing GFP-Nup88 (full-length) or GFP vector alone. At 48 h after transfection, the cells were fixed, stained with an anti-Nup88 antibody (red) and analyzed by confocal laser microscopy. Chromatin was stained with DAPI (blue). Scale bar, 10 μm. The white arrow indicates a typical multinucleated cell. (C) Representative images of mitotic HeLa cells transfected with the plasmid overexpressing GFP-Nup88 (full-length). The cells were fixed, stained with anti-Nup214 (red) antibody and analyzed by confocal laser microscopy. Chromatin was stained with DAPI (blue). Scale bar, 10 μm. The white arrow indicates a typical multinucleated cell. (D) Mitotic HeLa cells were scored for multinucleated cell defects. The data represent the means of three experiments in which 250 mitotic cells were scored in three independent experiments.