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. 2014 Jan 1;25(1):30–40. doi: 10.1091/mbc.E13-08-0479

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© 2014 Kim and Smith. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 3: — TIN2C or SA1 overexpression leads to anaphase delay. (A) Schematic diagram depicting persistent telomere cohesion and loss of centromere cohesion induced by TIN2C or SA1 overexpression. (B, C, G, H) TIN2C and SA1 overexpression leads to a loss in centromere cohesion. HTC75 cells stably expressing (B, C) V or TIN2C or (G, H) V or SA1 were analyzed by (B, G) centromere (red) and telomere (green) FISH after mitotic shake-off. DNA was stained with DAPI (blue). Scale bars, 5 μm. (C, H) Graphical representation of the frequency of mitotic cells with centromeres apart and telomeres cohered. Values are means ± SEM, derived from two independent experiments (C, n = 50–56 cells each; H, n = 50–60 cells each). (D–F, I–K) Live-cell imaging indicates that TIN2C or SA1 overexpression induces anaphase delay. Time-lapse video live-cell imaging of (D–F) HTC75.Vector-H2B-GFP and HTC75.TIN2C-H2B-GFP or (I–K) HTC75.Vector-H2B-GFP and HTC75.SA1-H2B-GFP cell lines. (D, I) Progression from prophase to anaphase for individual cells. Scale bars, 5 μm. (E, F, J, K) Graphical summaries of individual mitotic cells (E, F; n = 49–56 cells each from two independent experiments) and (I–K; n = 18–21 cells each) shown as (E, J) a time line and (F, K) a scatterplot with calculated mean value ± SEM. Student's t test was used to calculate p values (***p ≤ 0.001; ****p ≤ 0.0001).