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Fig. 6 | Cell Division

Fig. 6

From: Mitotic arrest affects clustering of tumor cells

Fig. 6

Metaphase-blocked MCF-7 cells do not form actin-dependent protrusions during anchorage-independent aggregation. a Representative fluorescence images of metaphase-synchronized/blocked MCF-7 cells at different time-points during clustering. The dotted line shows the micro-well edge. Scale bar: 50 µm. b Left: Images of a PDMS micro-well with untreated cells (top) and metaphase-synchronized/blocked MCF-7 cells (Met-sync, bottom) at the last time point of the time-lapse experiment (180 min). Right: Corresponding binary images at the end of the image segmentation process. Circles, edge of the micro-well; red, the largest cluster formed that was used for the aspect ratio and circularity analysis. c Graphs showing the aspect ratio (left) and circularity (right) analysis results for the larger clusters in control (untreated; UNT) and metaphase-synchronized (Met-sync) cells after 3 h of clustering. Each dot corresponds to the values in one micro-well from 5 independent experiments and bars correspond to the mean ± SD. *P < 0.05; **P < 0.01 (Mann–Whitney non-parametric test). d Representative micrographs of control (untreated) and metaphase-synchronized/blocked (Met-sync) MCF-7 cells that express the LifeAct-mCherry fluorescent reporter during aggregation (Left panels). The white line shows the region of interest (ROI) used for the aspect ratio determination. e Graphs showing the average (µ) and standard deviation (σ) aspect ratio of control (UNT) and in metaphase-synchronized/blocked cells (Met-sync) after 1 h of aggregation. Each dot corresponds to one cell and the bars correspond to the mean ± SD. Data are from 5 independent experiments with 5-6 cells analyzed per experiment. ****P < 0.0001 (Mann–Whitney non-parametric test)

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