Figure 1.
NE defects appear in a cell cycle-specific manner in Torsin-deficient cells. (A) EM images of NE herniations in Torsin-deficient (4TorKO) HeLa cells. EM images of mature NPCs from WT cells are shown for comparison. Black arrow, mature NPCs; white arrow, NE herniation; N, nucleus; C, cytoplasm. Scale bars, 250 nm. (B) Graph comparing the number of mature pores and NE herniations with visible neck regions found in WT and 4TorKO cells. Quantifications are based on the number of mature pores or attached herniations observed per 30 µm of NE with each point on the graph representing an individual cross section. The average number of blebs per 30 µm of NE and the standard error of the mean are indicated by the bar graph and error bars, respectively. 30 cells were counted for each sample. (C) Diagram depicting the cell cycle stages in HeLa cells (∼20 h per complete cycle). In synchronization experiments, cells were arrested at the G1 to S phase transition (red arrowhead), which was designated as the 0 h time point (T = 0 h). In D–F, the times given are the amount of time after release from the double thymidine block. (D) Confocal images of three different time points from synchronized 4TorKO cells stained with a K48-Ub antibody. White arrow: midbodies. Scale bar, 10 µm. (E) Graph showing the percent of cells exhibiting K48-Ub foci at eight different time points from synchronized cells and an asynchronous control. Mitosis occurs between the 7- and 12-h time points (dashed red line). Each time point shows the average of three independent experiments with 50 cells each. Error bars indicate ±SD. (F) Graph of the number of blebs observed per EM cross section in samples from synchronized cells at two time points or asynchronous cells. Each point on the graph represents the number of blebs per 30 µm of NE in an individual cross section. The average number of blebs per 30 µm of NE and the standard error of the mean are indicated by the bar graph and error bars, respectively. 30 cells were counted for each sample. P value was calculated using a two-tailed t test.

NE defects appear in a cell cycle-specific manner in Torsin-deficient cells. (A) EM images of NE herniations in Torsin-deficient (4TorKO) HeLa cells. EM images of mature NPCs from WT cells are shown for comparison. Black arrow, mature NPCs; white arrow, NE herniation; N, nucleus; C, cytoplasm. Scale bars, 250 nm. (B) Graph comparing the number of mature pores and NE herniations with visible neck regions found in WT and 4TorKO cells. Quantifications are based on the number of mature pores or attached herniations observed per 30 µm of NE with each point on the graph representing an individual cross section. The average number of blebs per 30 µm of NE and the standard error of the mean are indicated by the bar graph and error bars, respectively. 30 cells were counted for each sample. (C) Diagram depicting the cell cycle stages in HeLa cells (∼20 h per complete cycle). In synchronization experiments, cells were arrested at the G1 to S phase transition (red arrowhead), which was designated as the 0 h time point (T = 0 h). In D–F, the times given are the amount of time after release from the double thymidine block. (D) Confocal images of three different time points from synchronized 4TorKO cells stained with a K48-Ub antibody. White arrow: midbodies. Scale bar, 10 µm. (E) Graph showing the percent of cells exhibiting K48-Ub foci at eight different time points from synchronized cells and an asynchronous control. Mitosis occurs between the 7- and 12-h time points (dashed red line). Each time point shows the average of three independent experiments with 50 cells each. Error bars indicate ±SD. (F) Graph of the number of blebs observed per EM cross section in samples from synchronized cells at two time points or asynchronous cells. Each point on the graph represents the number of blebs per 30 µm of NE in an individual cross section. The average number of blebs per 30 µm of NE and the standard error of the mean are indicated by the bar graph and error bars, respectively. 30 cells were counted for each sample. P value was calculated using a two-tailed t test.

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