Figure 7.
Figure 7. NPC clustering impairs nuclear import and export of GFP-reporter proteins. (A) Image series showing import of NLS-GFP after photobleaching. Images were subject to adjustment in brightness, contrast, and gamma settings. Time is given in minutes, the bar represents micrometers. (B) Graph showing fluorescent recovery of NLS-GFP in the nucleus after photobleaching in untreated cells and in the presence of the solvent DMSO. Data points represent mean ± SEM (n = 10). (C) Bar chart showing fluorescent recovery of NLS-GFP in control cells at 10 min after photobleaching in motor mutants and after 10 min of benomyl treatment, but before NPC clustering is visible. Clustering of nuclear pores due to deleting or inactivation of motors significantly affects import, whereas disruption of microtubules before clustering of NPCs has no significant effect on NLS-GFP import. Bars represent mean ± SEM (n = 31–58 per bar). **, significant difference to control at P < 0.01; ***, P < 0.0001. (D) Image series showing export of NLS-NES-paGFP after photoactivation. Decay of the export reporter is shown in false-colored images, where signal intensity is given in colors. Images were subject to adjustment in brightness, contrast, and gamma settings. Time is given in minutes, the bar represents micrometers. (E) Graph showing decay of fluorescent NLS-NES-paGFP after photoactivation in the nucleus in untreated cells and in the presence of the solvent DMSO. Data points represent mean ± SEM (n = 10). (F) Bar chart showing decrease of NLS-NES-paGFP in control cells, motor mutants, and after 10 min of benomyl treatment in cells that do not show clustering of the NPCs. Clustering of nuclear pores due to deleting or inactivation of motors slows down export of the reporter, whereas disruption of microtubules before clustering of NPCs has no significant effect on NLS-NES-paGFP export. Bars represent mean ± SEM (n = 31–40 per bar). ***, significant difference to control at P < 0.0001.

NPC clustering impairs nuclear import and export of GFP-reporter proteins. (A) Image series showing import of NLS-GFP after photobleaching. Images were subject to adjustment in brightness, contrast, and gamma settings. Time is given in minutes, the bar represents micrometers. (B) Graph showing fluorescent recovery of NLS-GFP in the nucleus after photobleaching in untreated cells and in the presence of the solvent DMSO. Data points represent mean ± SEM (n = 10). (C) Bar chart showing fluorescent recovery of NLS-GFP in control cells at 10 min after photobleaching in motor mutants and after 10 min of benomyl treatment, but before NPC clustering is visible. Clustering of nuclear pores due to deleting or inactivation of motors significantly affects import, whereas disruption of microtubules before clustering of NPCs has no significant effect on NLS-GFP import. Bars represent mean ± SEM (n = 31–58 per bar). **, significant difference to control at P < 0.01; ***, P < 0.0001. (D) Image series showing export of NLS-NES-paGFP after photoactivation. Decay of the export reporter is shown in false-colored images, where signal intensity is given in colors. Images were subject to adjustment in brightness, contrast, and gamma settings. Time is given in minutes, the bar represents micrometers. (E) Graph showing decay of fluorescent NLS-NES-paGFP after photoactivation in the nucleus in untreated cells and in the presence of the solvent DMSO. Data points represent mean ± SEM (n = 10). (F) Bar chart showing decrease of NLS-NES-paGFP in control cells, motor mutants, and after 10 min of benomyl treatment in cells that do not show clustering of the NPCs. Clustering of nuclear pores due to deleting or inactivation of motors slows down export of the reporter, whereas disruption of microtubules before clustering of NPCs has no significant effect on NLS-NES-paGFP export. Bars represent mean ± SEM (n = 31–40 per bar). ***, significant difference to control at P < 0.0001.

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