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Results: 5

1.
Figure 4

Figure 4. From: Formation of the Nuclear Envelope Permeability Barrier Studied by Sequential Photoswitching and Flux Analysis.

Equilibration rates decrease gradually during the first two hours after mitosis. (A) Equilibration rates for repeated permeability measurements after mitosis in 13 cells. The data points of three individual cells are highlighted (crosses for the cell shown in Fig. 2). (B) Close-up of the highlighted region in A. Data points displayed as empty circles show the average of all 13 cells. Error bars represent the standard deviation.

Elisa Dultz, et al. Biophys J. 2009 October 7;97(7):1891-1897.
2.
Figure 3

Figure 3. From: Formation of the Nuclear Envelope Permeability Barrier Studied by Sequential Photoswitching and Flux Analysis.

Nuclear envelope permeability is increased after mitosis. (A) Nuclear envelope permeability was probed repeatedly in the same cell during the first 1.5 h after mitosis. Dronpa was activated in the cytoplasm in the area indicated by a blue circle. (B) Mean nuclear fluorescence corrected for acquisition photobleaching is plotted. Curves were fitted with single exponentials. The fitted equilibration rate, s, and its standard error are given in C (see also Fig. 3). AO, anaphase onset.

Elisa Dultz, et al. Biophys J. 2009 October 7;97(7):1891-1897.
3.
Figure 1

Figure 1. From: Formation of the Nuclear Envelope Permeability Barrier Studied by Sequential Photoswitching and Flux Analysis.

Assay for measuring nuclear envelope permeability in living cells. An interphase NRK cell expressing soluble Dronpa was reversibly photoswitched by 5 s of illumination with a mercury lamp via a YFP filter. Subsequently, the area indicated by a dashed circle was activated with a brief pulse of 405 nm laser light. Redistribution of the activated Dronpa was monitored. Nuclear fluorescence was quantified (dashed white outline), corrected for acquisition photobleaching with whole-cell fluorescence (white outline), and fitted with a single exponential (black solid line). Scale bar, 10 μm.

Elisa Dultz, et al. Biophys J. 2009 October 7;97(7):1891-1897.
4.
Figure 2

Figure 2. From: Formation of the Nuclear Envelope Permeability Barrier Studied by Sequential Photoswitching and Flux Analysis.

Kinetics of IBB import into the nucleus. NRK cells stably expressing IBB-DiHcRed were imaged at ∼30-s time resolution through mitosis. IBB intensity in the nucleus was quantified in a manually segmented region. The quantification results of five different cells were normalized to a lower and upper plateau, aligned along the time point of anaphase onset, and averaged. Error bars show the standard deviation. Mean time from anaphase onset to half-maximal intensity in the nucleus was 6.8 ± 0.5 min. Images were filtered with a Gaussian blur filter (kernel size 4) for presentation purposes.

Elisa Dultz, et al. Biophys J. 2009 October 7;97(7):1891-1897.
5.
Figure 5

Figure 5. From: Formation of the Nuclear Envelope Permeability Barrier Studied by Sequential Photoswitching and Flux Analysis.

Models for increased NPC permeability after mitosis. (A) Although NPCs already contain most subunits shortly after mitosis, the addition of components continues into G1 and could be required to establish the full permeability barrier. (B) The assembly of new NPCs may occur at an increased rate early after mitosis. Early assembly intermediates probably include stages where the inner and outer nuclear membranes have already fused but the pore is not yet occupied by NPC proteins. (C) During mitosis, the transcription of RNAs is largely shut down, so that at early time points after mitosis, most of the large and abundant cargoes transported through the NPC in interphase are not present. This reduced transport load might lead to a higher permeability of the NPC for passive diffusion.

Elisa Dultz, et al. Biophys J. 2009 October 7;97(7):1891-1897.

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