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Fig. 1. Localization of NE proteins on the telophase chromosome. (A) Schematic diagram of the `core' region of NE proteins on telophase chromosomes. The region marked in red represents the core region where BAF is initially accumulated. The region marked in green represents the non-core region outside the core region. (B) Living HeLa cells fluorescently stained with Hoechst 33342 for chromosomes and with GFP fusions of NE proteins (as indicated on the left) were observed every minute. The time-lapse images obtained are shown from left to right, with time 0 indicating the metaphase-anaphase transition. BAF, lamin A and all LEM domain proteins are localized at the `core' region. Arrows indicate the time of core localization obtained from quantification of core localization shown in C. Scale bar: 10 µm. (C) Fluorescence intensity at the core region is plotted as a function of time in minutes after the metaphase-anaphase transition. Fluorescence intensity is presented as % maximum intensity after subtracting the intensity at 4 minutes. (D) The time of core localization was estimated from the plots shown in C. The time of core localization is defined by the time required, in seconds after the metaphase-anaphase transition, for intensity at the core to reach 80% of the maximum. The s.d. and the number of cells tested (in parentheses) is also shown. (E) Schematic diagram of NE formation. BAF (red) localizes first to the core region, prior to other LEM domain proteins.
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