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. 2021 Jan 27;12:608. doi: 10.1038/s41467-020-20801-0

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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 Workflow: aged chimeras, analysed in (b–d). b Absolute frequencies of bone marrow CD45.2⁺ HSPCs in full Wt and Neo1^gt//gt chimeras after 15 months; n = 7 (Ctrl)–11 (Neo1), two independent experiments. c Absolute blood counts of full Wt and Neo1^gt//gt chimeras after 15 months; n = 7 (Ctrl)–11 (Neo1), two independent experiments, for HB: 4 (Ctrl)–7 (Neo1). d Frequencies of B cells and myeloid cells of C45.2⁺ cells in peripheral blood of Wt and Neo1^gt//gt chimeras after 15 months; n = 5–13 (for exact n/timepoint please see Source data file), two independent experiments. Analysis with two-way-ANOVA, multiple comparisons with LSD Fisher’s test. e Workflow: assessment of HSC potency derived from 15 months (aged) chimeras. f Frequency of CD45.2⁺ vs. competitor HSCs 16 weeks transplantation of 100 or 500 HSCs from of aged Wt and Neo1^gt//gt chimeras; n = 6 (Ctrl + 500 HSC Neo1)–7(100 HSC, Neo1), two independent experiments. g Workflow: secondary and tertiary transplantations of 15 months (aged) chimeras. h Absolute frequencies of bone marrow CD45.2⁺ HSPCs in 2° transplantations of aged Wt and Neo1^gt//gt chimeras after 4 months; n = 7 (Ctrl)–8 (Neo1), two independent experiments. i Absolute frequencies of bone marrow CD45.2⁺ HSPCs in 3° transplantations of aged Wt and Neo1^gt//gt chimeras after 4 months; n = 6, two independent experiments. j Workflow: generation of full chimeras used in (k–m). k Cell cycle phase of CD45.2⁺ HSCs derived from Wt and Neo1^gt//gt chimeras after 4 months; n = 4 (Ctrl)–6 (Neo1), two independent experiments. l MFI of CDK6 in CD45.2⁺ HSC derived from Wt and Neo1^gt//gt chimeras after 4 months; n = 23 (Neo1)–29 (Ctrl). m Frequency of BrdU⁺ CD45.2⁺ HSC derived from Wt and Neo1^gt//gt chimeras after 4 months, 48 h post BrdU injection; n = 6, two independent experiments. For all panels, ±SD is shown. n indicates biological replicates. P value was determined by two-tailed t test unless stated otherwise. Scale bars in IF images are 5 μm. Source data are provided as a Source Data file.