Figure 5.
Figure 5. Disassembly kinetics of F-actin patches are tightly correlated with chromosome capture by microtubules. (A) Maximum-intensity z-projection of selected time points from a time series of the nuclear area of an oocyte expressing mEGFP-ArpC1 to label the Arp2/3 complex (gray), and H2B-mCherry to label chromosomes (cyan). Last frame: Chromosome tracks overlaid, color-coded as below. See also Video 5. Below: Single confocal slices of selected chromosomes marked by dashed rectangles on the overview. Bars: (top) 10 µm; (bottom) 5 µm. Time is given as minutes:seconds relative to NEBD. (B) Plot of chromosome distance to the AP over time, calculated from the trajectories shown in A. Trajectories are color-coded for actin- (green) and microtubule (red)-driven transport phases and arrival at the spindle (gray). Chromosome capture events are represented by black dots. (C) Normalized mEGFP-ArpC1 intensity profile for each chromosome tracked in B. Intensity is calculated in a 5-µm diameter sphere around the chromosome’s center of mass and normalized to the background level before NEBD onset (in gray). Plots are color-coded as in B. (D) Individual plots for chromosomes shown in A. (E) Oocytes expressing 3mCherry-UtrCH to label F-actin (gray) and H2B-3mEGFP to label chromosomes (cyan) were then treated with DMSO or Latrunculin B, respectively (left and right panels). Left side: Maximum z-projection of the first time point (00:30). Right side: Single confocal slices of selected chromosomes and an area sampling the F-actin network as marked by dashed rectangles on the overview. Below: Individual plots of mean intensities for chromosome patches (a and b) or network (c) shown above. Red bars indicate the chromosome capture event. Bars: (main images) 20 µm; (smaller images) 5 µm. Time is given as minutes:seconds relative to NEBD. (F) Plot of chromosome distance to the AP over time for the oocytes shown in E. Top right: Normalized 3mCherry-UtrCH intensity profiles for the 5-µm-diameter sphere surrounding each chromosome. Bottom right: Normalized mean F-actin network intensity profile.

Disassembly kinetics of F-actin patches are tightly correlated with chromosome capture by microtubules. (A) Maximum-intensity z-projection of selected time points from a time series of the nuclear area of an oocyte expressing mEGFP-ArpC1 to label the Arp2/3 complex (gray), and H2B-mCherry to label chromosomes (cyan). Last frame: Chromosome tracks overlaid, color-coded as below. See also Video 5. Below: Single confocal slices of selected chromosomes marked by dashed rectangles on the overview. Bars: (top) 10 µm; (bottom) 5 µm. Time is given as minutes:seconds relative to NEBD. (B) Plot of chromosome distance to the AP over time, calculated from the trajectories shown in A. Trajectories are color-coded for actin- (green) and microtubule (red)-driven transport phases and arrival at the spindle (gray). Chromosome capture events are represented by black dots. (C) Normalized mEGFP-ArpC1 intensity profile for each chromosome tracked in B. Intensity is calculated in a 5-µm diameter sphere around the chromosome’s center of mass and normalized to the background level before NEBD onset (in gray). Plots are color-coded as in B. (D) Individual plots for chromosomes shown in A. (E) Oocytes expressing 3mCherry-UtrCH to label F-actin (gray) and H2B-3mEGFP to label chromosomes (cyan) were then treated with DMSO or Latrunculin B, respectively (left and right panels). Left side: Maximum z-projection of the first time point (00:30). Right side: Single confocal slices of selected chromosomes and an area sampling the F-actin network as marked by dashed rectangles on the overview. Below: Individual plots of mean intensities for chromosome patches (a and b) or network (c) shown above. Red bars indicate the chromosome capture event. Bars: (main images) 20 µm; (smaller images) 5 µm. Time is given as minutes:seconds relative to NEBD. (F) Plot of chromosome distance to the AP over time for the oocytes shown in E. Top right: Normalized 3mCherry-UtrCH intensity profiles for the 5-µm-diameter sphere surrounding each chromosome. Bottom right: Normalized mean F-actin network intensity profile.

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