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. 2020 May 7;106(4):1000–1007. doi: 10.3324/haematol.2019.235150

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Copyright© 2021 Ferrata Storti Foundation

This article is distributed under the terms of the Creative Commons Attribution Noncommercial License (by-nc 4.0) which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.

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Figure 2. — The ASXL1^G643W variant has minimal impact on normal hematopoiesis. (A) Fluorescence-activated cell sorting (FACS)-based analysis of the peripheral blood of 6-month old Asxl1^+/+, Asxl1^G643W/+ and Asxl1^G643W/G643W mice (n>11 mice in each experimental group). The relative distribution of B cells (B220), T cells (CD3), neutrophilic granulocytes (Mac1-Gr1) and other monocytic/granulocytic cells (Mac1) was analyzed. (B) FACS-based analysis of the peripheral blood of 18-month old Asxl1^+/+, Asxl1^G643W/+ and Asxl1^G643W/G643W mice (n>5 mice in each experimental group). The relative distribution of B cells (B220), T cells (CD3), neutrophilic granulocytes (Mac1-Gr1) and other monocytic/granulocytic cells (Mac1) was analyzed. (C) Representative FACS profiles of the data from (A-B) with the gating strategy indicated. The FACS profile represents an Asxl1^+/+ control mouse. See the Online Supplementary Figure S1 for representative FACS plots of Asxl1^G643W/+ and Asxl1^G643W/G643W animals. (D) FACS-based analysis of bone marrow hematopoietic stem cell (HSC) and multipotent progenitor (MPP) subsets in 6-month old Asxl1^+/+, Asxl1^G643W/+ and Asxl1^G643W/G643W mice (n>5 mice in each experimental group). (E) Representative FACS profiles of the data from (D) with the gating strategy indicated. The FACS profile represents an Asxl1^+/+ control mouse. See the Online Supplementary Figure S1 for representative FACS plots of Asxl1^G643W/+ and Asxl1^G643W/G643W animals. (F) Lineage distribution of mature bone marrow (BM) subsets in 6-month old Asxl1^+/+, Asxl1^G643W/+ and Asxl1^G643W/G643W mice (n>5 mice in each experimental group). The relative distribution of B cells (B220), T cells (CD3), neutrophilic granulocytes (Mac1-Gr1) and other monocytic/granulocytic cells (Mac1) was analyzed. (G) Spleen weights of 18-month old Asxl1^+/+, Asxl1^G643W/+ and Asxl1^G643W/G643W mice (n>5 mice in each experimental group). (H) Competitive BM transplantation of 1:1 mixtures of CD45.2 donor BM cells from 6-month old Asxl1^+/+, Asxl1^G643W/+or Asxl1^G643W/G643W mice and CD45.1 competitor cells into lethally irradiated CD45.1 recipient mice. The ratio of CD45.2 to CD45.1 in peripheral blood is depicted (n>7 mice in each experimental group).