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. 2019 Sep 26;105(7):1857–1867. doi: 10.3324/haematol.2019.219188

Figure 2.

Figure 2

Expression of iNUP98-KMT2A results in LSK cell expansion with a competitive repopulation advantage. (A) Lineage negative (Lin−) bone marrow (BM) cells from pre-leukemic iNUP98-KMT2A mice and wildtype (WT) littermate controls (CTRL) were stained for c-Kit and Sca-1 and analyzed by flow cytometry. *P<0.05, unpaired t-test. (B) Lin− BM from pre-leukemic iNUP98-KMT2A mice and WT littermate controls were stained for markers of long-term hematopoietic stem cells (LT-HSC) (LSK, CD150+, CD48−) and multipotent progenitors (MPP) (LSK, CD34+, CD48+, CD150−) and analyzed by flow cytometry. The percentages of LSK identified as LT-HSC and MPP are shown. (C) Cell cycle analysis of LSK from pre-leukemic iNUP98-KMT2A mice and WT littermate controls. **P<0.01, unpaired t-test. (D) Competitive repopulation assay: lethally-irradiated CD45.1 WT recipients were transplanted with a 1:1 mixture of total BM cells from CD45.2 iNUP98-KMT2A and CD45.1 WT mice in the presence of doxycycline (DOX). The percentage of CD45.2 cells present in the peripheral blood was measured by flow cytometry over a period of 25 weeks. *P<0.05, unpaired t-test. (E) At week 25 of the competitive repopulation assay, the CD45.2+ cells were analyzed for percentages of Gr-1, CD3, and B220 markers. (F) Cellular BM chimerism of WT CD45.1+ mice 25 weeks after competitive transplantation with CD45.2+ iNUP98-KMT2A BM. All mice were exposed to DOX throughout the experiment.