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. 2021 May 4;12:2522. doi: 10.1038/s41467-021-22749-1

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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 scheme for deleting Maea in adult mice using Mx1-Cre. (The mouse symbol in these figures were modified and recreated from Servier Medical Art (https://smart.servier.com/), an open source of medical images). b Quantification of bone marrow (BM) HSC numbers at indicated time points after poly I:C injection (day 0: n = 4; days 7 and 21: n = 5; day 14: n = 3 each group). Data are represented as floating boxes with boundaries indicate minima to maxima of each dataset and middle line indicates the mean. Day 0 p = 0.069, day 7 p = 0.0457, day 14 p = 0.168, day 21 p = 0.005. c Representative FACS plots and cell cycle profiles of control (Ctrl) and Maea^Mx1-Cre (CKO) HSCs at 21 days after 1st poly I:C injection (n = 4). p = 0.0002. d Experimental scheme for deleting Maea in 1:1 wild-type (WT) and Maea^Mx1-Cre BM chimeras after stable (8 weeks) reconstitution. e Donor chimaerism in BM LSK and HSCs and peripheral blood total leucocytes from control and Maea^Mx1-Cre mixed chimeric mice at indicated time points after poly I:C injection (n = 10 over two independent experiments). f RNA-seq and Gene Set Enrichment Analysis (GSEA) of HSCs from BM of control and Maea^Csf1r-Cre young adults at 7–12 weeks of age. Three replicates with 2000 HSCs pooled from two mice each replicate were processed and analysed for each group. Top KEGG (Kyoto Encyclopaedia of Genes and Genomes) pathways that are up-regulated and down-regulated in Maea^Csf1r-Cre HSCs are shown. g Examples of GSEA enrichment plots showing enrichment of Proteasome, Oxidative phosphorylation and mTOR signalling pathways in Maea^Csf1r-Cre HSCs. h GSEA enrichment plot showing significant downregulation of lymphoid potential related gene set in Maea^Csf1r-Cre HSCs. i Heat map showing mean expression of HSC related genes in control and Maea^Csf1r-Cre HSCs. j Experimental scheme and quantification of LSKs and HSCs in control and Maea^Mx1-Cre mice treated with vehicle (Veh), carfilzomib (CFZ), rapamycin (Rapa) or N-acetylcysteine (NAC) for 3 weeks after poly I:C induction (Veh: n = 5, CFZ: n = 6, Rapa: n = 7, NAC: n = 3 over two independent experiments). k Donor chimaerism in peripheral blood of CD45.1 lethally irradiated wild-type recipients at indicated time points after competitive BM transplantation (BMT) of equal number of CD45.1 WT competitor BM cells and CD45.2 donor BM cells from indicated groups (n = 5 each group). All data are mean ± sem unless otherwise indicated. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 by unpaired two-sided t-test with 95% confidence level. ns not significant.