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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. 3 — a TRF analysis of HCT116 wild type and MCM10^+/- cells. Estimated PDs and location of peak intensity (yellow ovals) are indicated. b Signal free-ends and fragile telomeres in wild type (blue) and MCM10^+/- cells (red). Significance was calculated using an unpaired, two-tailed student’s t-test with ***<.001; n = metaphases analyzed. Scale bars are 1 µm. c TRAP assay from wild type and MCM10^+/- cells. The internal PCR control at 36 bp and telomerase products above 50 bp are noted. d TRF analysis of wild type and independent late passage MCM10^+/- cell populations (PD > 75). The location of peak intensity (yellow ovals) is indicated. e Genotyping of wild type (middle) or mutant alleles carrying a loxP site 3′ of exon 14 (upper) or a loxP scar (lower). A faint non-specific band is noted (asterisk). f Genotyping in populations analyzed in Fig. 3d showing alleles with one 3′ loxP site (upper) or a loxP scar (lower), and reverted alleles that retained or lost the 3′ loxP site. A faint non-specific band is noted (asterisk). g TRF analysis (top) and genotyping (bottom) in wild type and MCM10^+/- cells. PDs and location of peak intensity (yellow ovals) are indicated. h Western blot for MCM10 with quantification normalized to tubulin, relative to the first lane is indicated. PDs for each cell line are noted. i Average proliferation rate in wild type, MCM10^+/- and reverted cells normalized to early passage wild type cells, n = 6. Individual data points are indicated (gray circles). j Average cell cycle distribution of wild type, MCM10^+/- and reverted cell lines, n = 4. Individual data points are indicated (gray circles). Percentage of each population in G1- (green), S- (purple), and G2/M-phase (gray) is indicated. Error bars in h and i indicate SD and significance was calculated using an unpaired, two-tailed student’s t-test with *<0.05; **<0.01, ***<0.001. Source data for panels a, b, c, d, e, f, g, h, i, and j, including relevant exact p-values, are provided in the Source Data file.