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. 2011 Jul 15;6(7):e22395. doi: 10.1371/journal.pone.0022395

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Gu et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) LNCAP cells were transfected with pcDNA-GFP-p44/WDR77 or pcDNA-GFP-NLS1. After expression of the transfected DNAs, the cells were fused to Cos 7 cells to form heterokaryons. After incubation for 6 hr, the cells were fixed and counterstained with TO-PRO 3, which differentiates the human and monkey nuclei within the heterokaryon (arrows identify the human nuclei). The panel marked Bright shows the phase-contrast image of the heterokaryons; the broken line highlights the cytoplasmic edge. (B) Cytoplasmic localization of f:GFP-NES1 and nuclear localization of f:GFP-NLS1 in PC3 cells. PC3 cells stably expressing f:GFP-NES1 (top panels) or f:GFP-NLS1 (bottom panels) were stained with TO-PRO 3 and observed under a confocal microscope. (C) Identification of polypeptides that were associated with NES1 or NLS1. The bands corresponding to f:GFP-NES1 and f:GFP-NLS1 are indicated by arrows. Polypeptides specifically associated with f:GFP-NLS1 or f:GFP-NES are indicated by numbers and stars. The standard protein markers (Bio-Rad) are shown in lane 1.