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. 2022 Jun 6;24(6):872–884. doi: 10.1038/s41556-022-00925-9

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© The Author(s) 2022

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, Fluorescence intensity of nuclear HK2 in haematopoietic cell populations from cord blood. A.u., arbitrary units; n = 183 cells examined from three biological samples. b, CD34⁺-enriched normal haematopoietic cells were transduced with NLS1–HK2 or control vector and injected into the right femur of NOD/SCID-GF mice (n = 4 (EV) and 5 (NLS1–HK2) mice). Engraftment of transduced cord blood cells in the left femur of mice was measured using flow cytometry 8 weeks after the injection. c, Cells from b were injected into secondary mice and the engraftment efficiency was measured 8 weeks later using flow cytometry (n = 5 mice per group). d, Confocal microscopy images of HK2 and Tom20 staining in bone marrow cells from Vav-NLS–HK2 mice and control littermate wild-type mice. Scale bar, 10 µm. Images are representative of n = 20 biologically independent samples. e, Percentage of lin⁻ckit⁺Sca⁺CD48⁻CD150⁺ cells in the bone marrow of the Vav-NLS–HK2 mice and their littermate controls (n = 12 (wild-type control) and 10 (Vav-NLS–HK2) mice). f, Percentage of granulocyte-monocyte and common myeloid progenitor (left), and megakaryocyte–erythroid progenitor (right) cells in the bone marrow of the Vav-NLS–HK2 mice and their littermate controls (n = 9 (wild-type control) and 6 (Vav-NLS–HK2) mice). g, Bone marrow cells from Vav-NLS–HK2 mice or their littermates (CD45.2⁺; donor) were co-transplanted with CD45.1⁺ bone marrow cells as competitors (1:1 ratio) into B6.SJL recipient mice (CD45.1⁺). Reconstitution units (CD45.2/CD45.1) were analysed in the peripheral blood of the chimaera mice over the specified period using flow cytometry (n = 10 mice per group). h, Reconstitution efficacy in the bone marrow from g was analysed at week 12 (n = 10 mice per group). b,c,e–h, Statistical analyses were performed using a two-tailed unpaired Student’s t-test. Data represent the mean ± s.e.m. MPP, multipotent progenitors; MLP, multilymphoid progenitors; CMP, common myeloid progenitors; GMP, granulocyte-monocyte progenitors; and MEP, megakaryocyte–erythroid progenitors.

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