XB-IMG-119664
Xenbase Image ID: 119664
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Figure 5. Recruitment of nucleoporins and lamin B1 during nuclear assembly. (A) In vivo localization of transiently expressed POM121-YFP3, EGFP3-Nup153, and ECFP–lamin B1 in metaphase NRK cells. Shown are confocal sections and DIC images centered on the metaphase plate. Note reticular pattern for POM121 and diffuse distribution of Nup153 and lamin B1. (B) 4-D sequence (10 slices every 2.5 μm acquired every 10–60 s depending on the dynamics of the cell cycle stage) of an NRK cell transiently expressing EGFP2-Nup153. Transparent projection of the four slices containing chromosomes (top) and DIC images highlighting the position of the chromosomes (bottom) are shown. Recruitment of Nup153 on the surface of chromosomes starts at ∼4 min after metaphase to anaphase transition. Time, h:mm:ss normalized to meta/anaphase transition equals 0. (C) 4-D double label sequence (six slices every 3 μm acquired every 30–120 s depending on the dynamics of the cell cycle stage) of an NRK cell transiently coexpressing POM121-YFP3 (top) and ECFP–lamin B1 (bottom). Shown are maximum intensity projections of the z-slices containing chromosomes. POM121 recruitment starts ∼3.5 min after meta/anaphase transition, whereas lamin B1 is only seen at 9 min (see D). Time, h:mm:ss normalized to meta/anaphase transition equals 0. (D) Plot of mean fluorescence intensity of the reforming NE for POM121 (green) and lamin B1 (black) shown in B. Before visible accumulation of lamin, nuclear rim areas were identified by POM121 localization. Data was background subtracted and normalized for bleaching during the time series. Lines are moving averages of two frames. Note the delay of ∼5 min between peak concentration/area for POM121 versus lamin B1. Bars: (A) 5 μm; (B and C) 10 μm. Online supplemental material (Fig. S2) is available at http://www.jcb.org/cgi/content/full/200101089/DC1. Image published in: Daigle N et al. (2001) Copyright © 2001, The Rockefeller University Press. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license Larger Image Printer Friendly View |