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. 2021 May 21;35(10):2948–2963. doi: 10.1038/s41375-021-01267-5

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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 Experimental setup scheme. Bone marrow (BM) was harvested from Bl6 mice, enriched for Lin⁻ Sca1⁺ cKit⁺ (LSK) cell markers by flow cytometry and cells were transduced with overexpression (OE) lentiviral vectors for Evl or miR-342 (and mock as a control). The influence of Evl and miR-342 OE on the hematopoietic differentiation capacity was assessed by lymphoid and myeloid colony-forming assays (CFC) in vitro or a CFC-Spleen (S) assay in vivo. Multilineage reconstitution and self-renewal were addressed by serial BM transplantation. b Relative number of colonies derived from EVL-, miR-342- or mock overexpressing LSK cells in pre-B-cell growth supporting semisolid medium (n = 3). c Relative number of colonies derived from EVL-, miR-342- or mock overexpressing LSK cells in erythroid and myeloid growth supporting semisolid medium (n = 3). d Weight of spleens in mg of mice transplanted with 14,400 LSK cells/mouse (group #1), 1440 LSK cells/mouse (group #2), and 530 LSK cells/mouse (group #3) transduced with miR-342 or mock overexpression vectors 13 days post-transplant. e Number of myelopoietic progenitor-derived colony-forming units (CFU) within the spleens of mice transplanted with LSK cells OE miR-342 or mCherry control 13 days post-transplant. f Frequency of donor-derived (CD45.1) and transduced (mCherry) cells within spleens of mice at day 13 after transplantation of miR-342 OE or mock LSK cells. g Frequency of myeloid (CD11b, Gr1) and lymphoid (CD3, CD45R) hematopoietic cells within the mCherry-positive donor cell fraction in spleens of mice at day 13 after transplantation of miR-342 OE or mock LSK cells. h Frequency of donor-derived eGFP/mCherry-positive peripheral B-cells in primary (1°) and i secondary (2°) recipient mice. j Frequency of donor-derived eGFP/mCherry-positive peripheral granulocytes in primary (1°) and k secondary (2°) recipient mice. *P < 0.05; **P < 0.01; ***P < 0.001; h–k (n = 6 mice/group; SEM).