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. 2021 Nov 25;12:6850. doi: 10.1038/s41467-021-27206-7

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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. 4 — a Numbers of telomere dysfunction–induced foci (TIF) per HSC. Bars represent the means ± S.E.M. of TIF per HSC (n = 35 pooled HSCs from 4 G0, 2 G5/G6, 6 G0 + OHT, and 4 G5/G6 + OHT-treated mice); each dot represents one HSC. Statistically significant differences were detected using one-way ANOVA. ***P < 0.001, ****P < 0.0001; G0 vs G0 + OHT: P = 0.72. b UMAP of scRNA-seq data displaying 1638, 1926, 1533, and 1777 pooled single HSCs isolated from R26-LSL G0 mice treated with vehicle (G0) or OHT (G0 + OHT) and from R26-LSL G5/G6 mice treated with vehicle (G5/G6) or OHT (G5/G6 + OHT), respectively (n ≥ 5 mice per group from two independent experiments of telomerase reactivation). Each dot represents one cell. Different colors represent the sample (left; dashed lines indicate the majority of G5/G6 HSCs) and cluster (right) identities. c Differences in the blood cell counts of each mouse between the start and end of treatment. Each dot represents one mouse (n = 15 G0, 14 G5/G6, 18 G0 + OHT, and 19 G5/G6 + OHT-treated mice from four independent experiments of telomerase reactivation). Data were expressed as percentages of delta variation ([cell blood count after treatment–cell blood count before treatment]/cell blood count before treatment). Bars represent the means ± SEM. Statistically significant differences were detected using one-way ANOVA. **P < 0.01, ***P < 0.001, ****P < 0.0001. Neutro neutrophils, Lympho lymphocytes, RBC red blood cells, PLT platelets. d Frequencies of CD45.2⁺ cells in the BM of CD45.1 recipients that were competitively transplanted with equal numbers of HSCs (n = 200) isolated from R26-LSL mice with the indicated genotypes and treatments (n = 9 G0, 5 G5/G6, 11 G0 + OHT, and 6 G5/G6 + OHT-treated mice from two independent experiments of telomerase reactivation). Bars represent the means ± SEM. Statistically significant differences were detected using one-way ANOVA. ****P < 0.0001; G0 vs G0 + OHT: P = 0.40. e UMAP of scRNA-seq data of pooled single HSCs isolated from G0 mice treated with a control oligonucleotide (G0 + control-ODN; 1021 HSCs) or the oligodeoxynucleotide A151 (G0 + A151-ODN; 1129 HSCs) and from G5/G6 mice treated with a control oligodeoxynucleotide (G5/G6 + control-ODN; 929 HSCs) or the oligodeoxynucleotide A151 (G5/G6 + A151-ODN; 955 HSCs) (n ≥3 mice per group). Each dot represents one cell. Different colors represent the sample (left) and cluster (right) identities. f Pathway enrichment analysis of significantly downregulated genes in A151-ODN–treated G5/G6 HSCs from cluster 2 shown in Fig. 4e, as compared to those in control-ODN–treated G5/G6 HSCs (adjusted P ≤ 0.05). The top ten Hallmark and Reactome gene sets are shown. Source data are provided as a Source Data file.