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. 2010 Jan-Feb;1(1):71–84. doi: 10.4161/nucl.1.1.10244

Figure 3.

Figure 3

Overexpression of the nuclear pore complex assembly region (NPAR) of Nup153 induces mitotic abnormalities. HeLa cells expressing histone H2B-GFP were transfected with GFP-Nup153 (A–C), GFP-Nup153-39-339 (D; i.e., the NPAR of Nup153), GFP-Nup153-39-144 (E) or GFP-Nup153-145-339 (F) and prepared for immunofluorescence 48 hours post transfection using monoclonal antibodies directed against β-tubulin (E and F). Cells expressing either GFP-Nup153, GFP-Nup153-39-339, or GFP-Nup153-145-339 showed poor spindle morphology and a strong increase in the frequency of multipolar spindles (A, B, D and E) and lagging chromosomes (C). Cells expressing the N-terminal portion of the NPAR, GFP-Nup153-39-144, show predominantly normal bipolar spindles (F). Scale bars, 5 µm. (G) Quantification of the number of multipolar spindles among mitotic cells. Cells were significantly more likely to have multipolar mitotic spindles when expressing Nup153 (26.2 ± 3.7%), Nup153-39-339 (24.7 ± 2.5%), or GFP-Nup153-145-339 (19.2 ± 5.5%) than control cells or cells expressing Nup153-39-144. Spindles were counted from 3–4 independent experiments with typically 100–200 spindles per experiment.