Figure 3. Plk4 depletion leads to a... | Rockefeller University Press

Figure 3.

$Figure 3. Plk4 depletion leads to a failure of centriole duplication followed by a cell cycle arrest. (A) Graph showing the fold increase in cell number after IAA addition. Plk4 destruction leads to a cell cycle arrest in Plk4AID/AID cells. Points show the mean of at least two independent experiments performed in triplicate. (B) Quantification of the number of Centrin foci per cell in interphase at indicated times after IAA addition. Bars represent the mean of >100 cells from two independent experiments. (C) Graph showing the fold increase in cell number after IAA washout (WO) and restoration of Plk4 levels. Centriole loss leads to an irreversible cell cycle arrest. Points show the mean of at least two independent experiments performed in triplicate. (D) Quantification of the duration of mitosis. Measurements were taken over a 24-h period at indicated times after IAA addition. Bars represent the mean of >60 cells from two independent experiments. (E) Quantification of relative cell proliferation and fraction of cells undergoing chromosome missegregation or cytokinesis failure. Measurements were taken over a 24-h period at indicated times after IAA addition. Bars represent the mean of >40 cells from two independent experiments. (F) Quantification of the frequency of divisions resulting in the formation of misshapen nuclei. Measurements were taken over a 24-h period at indicated times after IAA addition. Bars represent the mean of >40 cells from two independent experiments. All error bars in the figure represent the SEM. (G) Selected images from a time-lapse series of untreated or IAA-treated Plk4AID/AID cells coexpressing histone TagRFP-tubulin, EGFP-Histone H2B, and EGFP-CEP63. Insets show EGFP-CEP63 at the centrosome. Time is indicated in minutes relative to nuclear envelope breakdown (time point 0). Note that the cell treated with IAA for 2 d exhibits an asymmetric spindle with one acentriolar pole. The interphase nucleus (asterisk) at 189 min later left the field of view. Bars: (main) 5 µm; (inset) 1 µm.$

Plk4 depletion leads to a failure of centriole duplication followed by a cell cycle arrest. (A) Graph showing the fold increase in cell number after IAA addition. Plk4 destruction leads to a cell cycle arrest in Plk4^AID/AID cells. Points show the mean of at least two independent experiments performed in triplicate. (B) Quantification of the number of Centrin foci per cell in interphase at indicated times after IAA addition. Bars represent the mean of >100 cells from two independent experiments. (C) Graph showing the fold increase in cell number after IAA washout (WO) and restoration of Plk4 levels. Centriole loss leads to an irreversible cell cycle arrest. Points show the mean of at least two independent experiments performed in triplicate. (D) Quantification of the duration of mitosis. Measurements were taken over a 24-h period at indicated times after IAA addition. Bars represent the mean of >60 cells from two independent experiments. (E) Quantification of relative cell proliferation and fraction of cells undergoing chromosome missegregation or cytokinesis failure. Measurements were taken over a 24-h period at indicated times after IAA addition. Bars represent the mean of >40 cells from two independent experiments. (F) Quantification of the frequency of divisions resulting in the formation of misshapen nuclei. Measurements were taken over a 24-h period at indicated times after IAA addition. Bars represent the mean of >40 cells from two independent experiments. All error bars in the figure represent the SEM. (G) Selected images from a time-lapse series of untreated or IAA-treated Plk4^AID/AID cells coexpressing histone TagRFP-tubulin, EGFP-Histone H2B, and EGFP-CEP63. Insets show EGFP-CEP63 at the centrosome. Time is indicated in minutes relative to nuclear envelope breakdown (time point 0). Note that the cell treated with IAA for 2 d exhibits an asymmetric spindle with one acentriolar pole. The interphase nucleus (asterisk) at 189 min later left the field of view. Bars: (main) 5 µm; (inset) 1 µm.