Fig. 5.

TOP2A depletion prevents prometaphase condensation. (A) Diagram of experimental conditions for B–F. TOP2A-mAID +H2B-EGFP cells were seeded in chambered slides and grown in media +/− auxin for 60 min before being imaged over time by spinning-disk confocal microscopy. (B) Example of an untreated TOP2A-mAID cell going through mitosis. (C) Example of an auxin-treated TOP2A-mAID cell entering and exiting mitosis. (B and C) Fluorescence intensity is displayed as fire LUT. (Scale bars: 5 μm.) (D) Quantification of the duration of mitotic phases from live cell imaging experiments. Each data point is an average of the n number of cells. The data were collected from at least three independent experiments. Error bars denote SEM. Asterisks denote P values from parametric Student t tests: ***P < 0.001. (E and F) Three-dimensional modeling analysis of chromatin volume (E) and density (F) of nuclei entering mitosis. Values are normalized to the time point 40 min before nuclear envelope breakdown (NEB), which is set as 1. The dashed lines mark NEB. Each data point is an average of n number of nuclei analyzed. Error bars denote SEM.