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Fig. 6. 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-c 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). (H) Schematic diagram of NE reformation during telophase. Upper panel: cartoon representation of NE formation at the core region. BAF (green) forms a stable complex on the surface of the telophase chromosome mass in the presence of MTs. Then, LEM-domain proteins containing NE precursor vesicles attach to the BAF complex, fuse with each other, and finally enclose the chromosomes at the core region as MTs disassemble. Lower panel: EM images of the stages depicted by the cartoon representations. Scale bars: 10 µm (A,C), 2 µm (D), 200 nm (G) and 100 nm (H).

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