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. 2021 Feb 16;12:1065. doi: 10.1038/s41467-021-21317-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. 2 — a Relative mRNA expression of Light (red), Lta (black), and Ltb (gray) by FACS-purified LSKs from naive mice (n = 3) and 1st chimeras 6 weeks post transplantation (n = 4 mice, two were pooled for the analysis). b Relative mRNA expression of Ltbr by FACS-purified LSKs from naive mice (n = 3 mice) and 1st chimeras 6 weeks after transplantation (n = 4 mice, two were pooled for the analysis). c Fold change in colony formation of Light^−/− BM LSKs (red) relative to BL/6 BM LSKs (black) in serial re-platings (n = 6). One out of two independent experiments is shown. d Percentage of CFU from Light^−/− (gray) or BL/6- GFP LSKs (black) in mixed colony-forming assays. The frequency of GFP⁺ (BL/6) vs. GFP⁻ (Light^−/−) colonies is shown. Data are pooled from two independent experiments and presented as mean ± SEM. e Experimental scheme for serial transplantations of Light^−/− Ly5.1 mixed BM chimeras. f–j Chimerism (red: Light^−/−; black: Ly5.1) in percentages of donor Lin⁻ cells (f), LT-HSCs (g), ST-HSCs (h), MPP1 (i), MPP2/3 (Lin⁻c-kit⁺sca1⁺CD150⁻CD48⁺) (j). Data are shown as mean ± SEM of n = 7 (1st), n = 6 (2nd), n = 3 (3rd, 4th) mice. k Experimental scheme of the serial transplantation of Ly5.1 and Light^−/− LSKs (1:1) into lethally irradiated Ly5.1/Ly5.2 and congenic Ly5.1/Light^−/− recipients. l–o Percentages of donor Lin⁻ cells in Ly5.1/Ly5.2 (l) and Ly5.1/Light^−/− recipients (m) and LSKs in Ly5.1/Ly5.2 (n), and Ly5.1/Light^−/− recipients (o). Data are shown as mean ± SEM, Ly5.1/Ly5.2 recipients: n = 7 (1st), n = 5 (2nd), n = 3 (3rd); n = 3 (4th) Ly5.1/Light^−/− recipients: n = 5 (1st), n = 5 (2nd), n = 3 (3rd), n = 3 (4th). One out of two independent experiments is shown, Statistics: *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 (two-tailed t test), a: p = 0.005; b: p < 0.0001; c: p = 0.0014; d: p = 0.014, f: 1^st p < 0.0001, 4th p < 0.0001; g: 1st p = 0.0289, 4th p < 0.0001; h: 1st p < 0.0001, 4th p < 0.0001; i: 1st p < 0.0001, 4th p < 0.0001; j: p < 0.0001, 4th p < 0.0001; l: 1st p < 0.0001, 4th p = 0.043; m: 1st p = 0.0004, 4th p < 0.0001; n: 1st p < 0.0001, 3rd p = 0.0005; o: p = 0.0005. .