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. 2020 Nov 9;40(2):436–451. doi: 10.1038/s41388-020-01524-4

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — a Model depicting the causal relationship between increased microtubule plus end growth rates in mitosis and the induction of chromosome missegregation constituting W-CIN. b Left panel: Model for the structure of p73 and DNp73: transactivation domain (TA), DNA-binding domain (DBD), oligomerization domain (OD), SAM domain (SAM), transactivation inhibitory domain (TID). Right panel: Doxycycline-inducible expression of DNp73 in chromosomally stable HCT116 cells. Three independent stable cell lines were treated with doxycycline for 24 h and expression of DNp73 was detected by western blotting. A representative western blot is shown. Note that the two bands for endogenous p73 represent the α and β isoforms of the protein. c Mitotic microtubule plus end assembly rates upon expression of DNp73. HCT116-DNp73 cell lines were treated with or without doxycycline and additionally with low-dose Taxol. As a control HCT116 parental cells were treated with doxycycline. Microtubule growth rates were determined by tracking EB3-GFP comets in mitotic cells by live-cell microscopy. Scatter dot plots show average microtubule polymerization rates (20 microtubules/cell, n = 30 mitotic cells from 3 independent experiments, mean ± SD, t-test). d Proportion of cells exhibiting lagging chromosomes in anaphase after expression of DNp73 and after restoration of proper microtubule growth rates. HCT116 parental cells were incubated with doxycycline as a control. Left: Representative examples of anaphase cells with or without lagging chromosomes (white arrows), Scale bar, 10 µm. Right: The graph shows the proportion of anaphase cells exhibiting lagging chromosomes for three independent stable cell lines (n = 300 anaphase cells from three independent experiments, mean ± SD, t-test). e Scheme depicting the generation of single-cell clones in order to determine chromosome number variability as a measure of W-CIN. f Proportion of cells harboring the indicated chromosome numbers per cell (n = 50 metaphase spreads). g Proportion of cells with chromosome numbers deviating from the modal (45 chromosomes in HCT116 cells). The calculation is based on analyses of chromosome number variability shown in f.