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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. 1 — a MCM10 schematic indicating NKD- (c.1276 C > T;c.1744C > T) and RCM-associated (c.236delG;c.764 + 5 G > A) variants and exons targeted using CRISPR-Cas9 (exon 3) or rAAV (exon14). b Pedigree and segregation of MCM10 variants in the RCM family. Blue shading indicates fetal RCM. Individuals that underwent exome (§) or genome (*) sequencing are indicated. Clinically unaffected children that are not carriers of both pathogenic variants (†) are indicated (carrier status of minors not disclosed). c Model of hMCM10-ID bound to single-stranded DNA, based on Xenopus laevis MCM10-ID (Protein Data Bank codes 3EBE¹⁶ [https://www.ncbi.nlm.nih.gov/Structure/pdb/3EBE] and 3H15⁷⁴ [https://www.ncbi.nlm.nih.gov/Structure/pdb/3H15]). Locations of the R426C (pink) and ΔN27 (red) variants and the zinc ion (gray sphere) are shown. d (Left) Coomassie blue-stained gel of WT and ΔN27 hMCM10-ID. (Right) Chromatography profiles of WT and ΔN27 hMCM10-ID. The molecular weight standard (gray) included thyroglobulin (670 kDa), γ-globulin (158 kDa), ovalbumin (44 kDa), myoglobin (17 kDa), and vitamin B12 (1.3 kDa). Trace profiles for WT and the molecular weight standard are offset on the y-axis for display purposes. e Western blot for MCM10 with GAPDH as a loading control. Quantification of MCM10 levels normalized to loading control, relative to wild type is indicated. f Average proliferation rate in MCM10^+/- cells normalized to wild type. For each cell line n = 6 replicates, with average values for biological replicates indicated (gray circles). g Comparison of clonogenic survival of HCT116 wild type (top) and MCM10^+/- cells (middle/bottom). Cells plated per well are noted. h Average percentage clonogenic survival in HCT116 wild type (blue) and MCM10^+/- cells (red), n = 15 replicates. Individual data points are indicated (gray circles). i Average percentage of early apoptotic, late apoptotic, or dead cells. Wild type (blue) and clonal MCM10^+/- cell lines (red) are shown with n = 4 biological replicates. Individual data points are indicated (gray circles). Error is indicated in f, h, and i as 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 d, e, f, h, and i, including relevant exact p-values, are provided in the Source Data file.