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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. 5 — a Mitotic microtubule polymerization rates upon induction of CDK1 activity mediated by inhibition of wee1. HCT116 cells were treated with increasing concentrations of the wee1 inhibitor MK-1775 in the absence or presence of the CDK1 inhibitor RO-3306 and mitotic microtubule plus end assembly rates were determined. Average microtubule polymerization rates are depicted in scatter dot plots (20 microtubules/cell, n = 30 mitotic cells from three independent experiments, mean ± SD, t-test). b Induction of lagging chromosomes upon CDK1 activation mediated by wee1 inhibition. HCT116 cells were treated with MK-1775 in the absence or presence of RO-3306. The graph shows the proportion of anaphase cells exhibiting lagging chromosomes (n = 300 anaphase cells from three independent experiments, mean ± SD, t-test). c Transient expression of FLAG-tagged CDK1-AF (constitutive active) or CDK1-DN (inactive) in chromosomally stable HCT116 cells. A representative western blot detecting CDK1 protein levels and exogenously expressed CDK1 (FLAG) is shown. d Mitotic microtubule polymerization rates after transient expression of CDK1-AF or CDK1-DN in HCT116 cells. Average microtubule polymerization rates are depicted in scatter dot plots (20 microtubules/cell, n = 30 mitotic cells from three independent experiments, mean ± SD, t-test). e Quantification of cells exhibiting lagging chromosomes after transient expression of CDK1-AF or CDK1-DN in the absence or presence of RO-3306 or Taxol (n = 300 anaphase cells from three independent experiments, mean ± SD, t-test). f Generation of HCT116 cell lines stably expressing CDK1-AF or CDK1-DN. The expression level of the CDK1 variants was detected in three independent cell clones. A representative western blot is shown. g Mitotic microtubule polymerization rates in single-cell clones of HCT116 cells stably expressing CDK1-AF or CDK1-DN. Average microtubule polymerization rates are depicted in scatter dot plots (20 microtubules/cell, n = 10 mitotic cells for each independent single-cell clone, mean ± SD, t-test). h Quantification of cells exhibiting lagging chromosomes in HCT116 single-cell clones stably expressing CDK1-AF or CDK1-DN (n = 300 anaphase cells for each independent single-cell clone, mean ± SD, t-test). i Chromosome number variability in single-cell clones derived from HCT116 cells stably expressing CDK1-AF or CDK1-DN. The proportion of cells with a chromosome number deviating from modal (45 chromosomes for HCT116 cells) was calculated for three independent clones (n = 50 cells per single-cell clone).