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V. Madan et al.
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Figure 3. ASXL2 deficiency perturbs frequency and function of hematopoietic stem cells (HSCs.) (A) Representative photograph of tibias and femurs, respectively, from >1-year old wild-type (WT) and Asxl2 knockout (KO) mice. (B) Bone marrow (BM) cellularity (two femurs and two tibias) in old WT and Asxl2 KO mice. (C) Proportion of LSK cells in the BM of old mice (n=14). (D) Frequency of BrdU+ LSK cells in the BM of 10-20-weeks old WT and Asxl2 deficient mice (n=5). (E) Proportions of CMP, GMP and MEP in the BM of >1-year old WT and KO mice (n=12). (F) Frequency of erythroid precursors (proE: CD71+TER119lo, EryA: CD71+TER119+FSChi, EryB: CD71+TER119+FSClo and EryC: CD71–TER119+FSClo) in BM of >1-year old WT and Asxl2 KO mice (n=8). (G-J) Competitive repopulation assay: 4000 (black circles) or 2000 (gray circles) LSK cells were injected in recipient mice and proportion of donor-derived cells (CD45.2+) in peripheral blood was determined at 4, 8, 12 and 16 weeks post transplantation. Ability of WT and Asxl2 KO LSK cells to reconstitute B cells (CD19+) (G), T cells (CD3+) (H), granulocytes (CD11b+Gr1+), (I) and monocytes (CD11b+Gr1–F4/80+) (J) was analyzed using flow cytometry. Bars represent mean±Standard Error of Mean. *P<0.05, ***P<0.001, ****P<0.0001, ns: not significant.
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haematologica | 2018; 103(12)