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. 2021 Dec 14;12:7268. doi: 10.1038/s41467-021-27197-5

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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 Example of IgM⁻IgD⁻ cell frequency evolution in a mouse that developed leukemia (top). Kaplan–Meier leukemia-free survival curves of control (Ctrl; n = 40) and IL7R mutant (IL7R^mut; n = 63) animals (bottom). All mice died with precursor B-ALL. Log-rank Mantel–Cox test (b) Histologies (H&E) of organs infiltrated with leukemia cells. Pie chart inserts represent fraction of analyzed animals (n = 4) with leukemia involvement (in black) in the respective organ. Scale bar indicates 100 μm (bone marrow, spleen), 250 μm (kidney, lung, liver), or 500 μm (brain). c Clonality pie charts based on IgH sequencing. Each colored slice corresponds to a clone, indicative of clonality. Gray areas correspond to many rare clones, indicative of polyclonality. Equitability values (ranging from 0, for monoclonality, to 1, for a balanced repertoire) are shown in the center of the pie charts. Samples are bone marrow pro+pre-B cells from control, pre-leukemic, or leukemic mice. d Ig heavy chain (H) over light chain (κ and λ) ratios to evaluate, at the population level, the presence of the pro-B cell rearrangement signature (heavy chain expression in the absence of light chain expression). Two-tailed Mann–Whitney test performed. e Principal component analysis of normal (Ctrl Pro+Pre-B) and pre-leukemic (Pre-leukemic) pro- and pre-B cell precursors, mature splenic B cells (Ctrl IgD⁺) and leukemia cells (IL7R^mut Leuk). f Immunophenotypic analysis of three representative BM leukemia samples. Numbers inside dot plots indicate frequency in each quadrant or region. g, h Kaplan–Meier leukemia-free survival curves of mice transplanted with g bulk primary leukemias or h sorted IgD⁺ versus IgM⁻IgD⁻ leukemia cells. i Myc and Bcl2 transcript upregulation (log2 fold change) in leukemia samples (n = 9) as compared to normal controls (n = 5). Moderated t-test performed. j Frequency of Ki67-positive cells in controls (n = 5), pre-leukemia (n = 5), and leukemia samples (n = 5). Dot plots are representative of each condition. k Frequency of annexin V/7AAD-negative (viable) cells in controls (n = 9) pre-leukemia (n = 8) and leukemia samples (n = 9). Dot plots are representative of each condition. j, k Two-tailed unpaired t-test. Source data are provided as a Source Data file.