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. 2018 Apr 17;8:6071. doi: 10.1038/s41598-018-24440-w

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© The Author(s) 2018

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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DNA repair and DDR gene expression, DSB marker and genome stability analyses. (A) left panel: relative expression of transcripts related to DNA repair, DDR checkpoint and apoptosis pathways in CPs (CMP, GMP and MEP) and mature blood-derived cells (B, T and NK) in comparison to HSPCs colored accordingly to the significance of the changes. right panel: color scheme description with blue colors for decreased expression and red colors indicative of increased expression relative to HSPCs along with the corresponding p values. (B) CD34⁺ cells were sorted into HSPC and CP fractions. Representative immunofluorescence staining visualized for 53BP1 (green) and DNA (blue) in HSPCs and CPs 6 hr after 3 Gy IR. (C) Average number of 53BP1 foci per cell in CPs and HSPCs. n = 3 experiments; Represented are mean values ± SD; *p < 0.05. At least 100 nuclei were analyzed in each experiment. (D) Workflow for the karyotype analysis of irradiated HSPCs and CPs. Freshly isolated CD34⁺ cells were sorted for HSPC and CP fractions, exposed or not to IR (2 Gy) and then plated at low density in methylcellulose to allow colony formation. Individual colonies were isolated for metaphase preparation and karyotype analysis (Table 1, and Supplementary Table 1).