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Figure 5


Fig. 5. Live-correlated EM analyses. (A) Time-lapse images of living HeLa cells expressing GFP-BAF were obtained every minute. Magenta represents chromosomes (Hoechst 33342) and green represents GFP-BAF. Cells were fixed after live-cell fluorescence imaging, and the same cell was subjected to EM analysis. (B) A series of deconvolved 3D images of the fixed cell. (C) A single section image from the indicated panel in B. (D) An EM image corresponding to the image in C. (E,F) Magnified views of the regions indicated in D. (G) High-magnification EM images of the regions a-d indicated in E and F. In the lower panels, drawings are superimposed on the EM images indicating GFP-BAF (green), the NE (red), MTs (orange) and vesicles (purple arrowheads). Scale bars: 10 µm (A,C), 2 µm (D) and 200 nm (G). (H) Time-lapse images of living HeLa cells obtained every minute. Upper panels represent chromosomes stained with Hoechst 33342 and lower panels fluorescently conjugated NLS-BSA microinjected as described by Haraguchi et al. (Haraguchi et al., 2000). Time 0 represents the metaphase-anaphase transition. Arrows in the image taken at 7 minutes indicate the first detectable accumulation of fluorescent NLS signals.





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