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. 2020 Oct 21;3:593. doi: 10.1038/s42003-020-01304-6

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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. 7 — a, b Comparison of average time spent in mitosis (a) and average time taken to complete mitosis (b) determined using time-lapse microscopy of the immortalized MEFs of indicated genotypes in the presence and absence of nocodazole (0.5 μM). Error bars represent the ± SD from two independent experiments (fate of n = 50 cell were counted per experiment). Student’s t test was performed to determine P-value < 0.05 (*), < 0.01 (**), < 0.001 (***) and < 0.0001 (****). c Percentage of mitotic outcome of the immortalized MEFs of indicated genotypes in presence of nocodazole (0.5 μM) as shown in panel A and B. Mitotic slippage was defined by premature mitotic exit during nocodazole-induced mitotic arrest, while death was determined through membrane blebbing. Mean derived from two independent experiments (fate of n = 50 cell were counted per experiment). d Cell-cycle profiles of immortalized MEFs of indicated genotype in the presence or absence of nocodazole (0.5 μM) determined using FACS. Error bars represent the ± SD from three independent experiments. Two-way ANOVA test was performed to determine P-value as demonstrated in Supplementary Table 3. e, f Polyploidy (>4N DNA content) (e) and percentage of SubG1 populations (f) determined using FACS in the indicated immortalized MEFs in the presence or absence of nocodazole (0.5 μM). Error bars represent the ± SD from three independent experiments with two replicates each. One-way ANOVA test was performed to determine P-value < 0.0001 (****). g Representative images of detryosinated (red) and acetylated tubulin (green) of metaphase stages of immortalized Cep55^wt/wt and Cep55^Tg/Tg MEFs showing the microtubule networks (Scale bar, 100 μm). h Quantification of the mean integrated density (MID) of Acetylated (Upper panel) and detyrosinated (lower panel) tubulin observed among immortalized Cep55^wt/wt and Cep55^Tg/Tg MEFs. The intensity was calculated using Image J software wherein n = 50 metaphase cells were calculated per genotype. Error bars represent the ± SD from two independent experiments. Student’s t test was performed to determine P-value < 0.0001 (****). i Percentage of mitotic defects (described previously in Fig. 6f upon KIF2B overexpression in the immortalized MEFs (n = 100 cell per experiment were counted) of indicated genotypes. Error bars represent the ± SD from three independent experiments. One-way ANOVA test was performed to determine P-value < 0.05 (*) and < 0.01 (**).