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. 2021 Mar 12;12:1626. doi: 10.1038/s41467-021-21878-x

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© The Author(s) 2021

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 Telomere combing images with telomeric DNA (green), nascent DNA (red), and merged images with examples of unreplicated telomeres (left), partially replicated (middle), and completely replicated telomeres (right). Telomeric and sub-telomeric regions are indicated. Scale bars are 3 µm. b Average percentage of unreplicated telomeres in wild type (blue) and MCM10^+/- cell lines (red) ST cell lines, n = total number of telomeres quantified. Average values for each replicate are indicated (gray circles). c Average percentage of completely replicated (yellow) versus partially/stalled telomeres (green) in ST cell lines, n = total number of replicated telomeres quantified. Average values for each replicate are indicated (gray circles). Error bars in b and c indicate SD and significance was calculated using an unpaired, two-tailed student’s t-test with *<0.05; **<0.01, ***<0.001. d (Left) Cartoon of t-complex DNA as formerly depicted³⁷. (Middle) Diagram of double-stranded telomere restriction fragment (ds-TRF), telomere circle (t-circle) and telomere complex (t-complex) DNA species from 2D gel electrophoresis. (Right) Comparison of 2D gels from ST cell lines. e Comparison of DNA species from 2D gel electrophoresis in HCT116 wild-type ST cells with and without 4-day HU treatment. f Comparison of DNA species from 2D gels in HCT116 wild type and MCM10^+/- ST cell lines with and without S1 nuclease digestion. g (Left) Image of t-SCE staining in ST cell lines. Examples of a t-SCE event and chromosomes without t-SCE are highlighted. (Right) Percentage t-SCE per chromosome ends. Bars represent average percentage t-SCE per chromosome ends; where n = the number of metaphases analyzed. Significance was calculated using an unpaired, two-tailed student’s t-test but were not statistically significant (n.s.). Scale bar is 10 µm. Source data for panels b, c, d, e, f, and g, including relevant exact p-values, are provided in the Source Data file.