Skip to main content
. 2020 Apr 24;4(8):1722–1736. doi: 10.1182/bloodadvances.2019000978

Figure 1.

Figure 1.

An EVI1 plus Gata2 High population has replicative potential. (A) Structures of the inv(3) allele, the 3q21q26-EVI1 BAC, and the 3q21q26-tdTomato BAC. In the 3q21q26-tdTomato BAC, a tdTomato reporter gene was introduced at the translation start site of the EVI1 gene in the parental 3q21q26-EVI1 BAC. (B) Dual-reporter mouse to monitor the effects of inv(3)-driven EVI1 and endogenous Gata2 expression by tdTomato and green fluorescent protein (GFP) fluorescence, respectively. (C) Representative flow cytometric profiles of bone marrow cells recovered from Gata2+/gfp::Tom or 3q21q26-EVI1::Gata2+/gfp::Tom mice. (D) Relative intensities of tdTomato (left) and GFP (right) fluorescence in High, Low, and Negative sorted populations (C). The average values for the High population in Gata2+/gfp::Tom mice was set as 1. (E) Relative mRNA levels of transgenic human plus endogenous mouse EVI1 (left) and mouse endogenous Gata2 mRNA (right). mRNA abundance was normalized to glyceraldehyde-3-phosphate dehydrogenase mRNA. The average values for the High population in Gata2+/gfp::Tom mice was set as 1. (F) Absolute number of High, Low, and Negative cells in Gata2 haploinsufficient mice either containing or lacking the 3q21q26-EVI1 BAC. (G) Percentages of “High” population cells in the bone marrows of Gata2+/gfp::Tom and 3q21q26-EVI1::Gata2+/gfp::Tom mice. (H) Colony-forming potential of High, Low, and Negative populations of Gata2+/gfp::Tom and 3q21q26-EVI1::Gata2+/gfp::Tom mice. Note that the EVI1-/Gata2-High population has high colony-forming potential, and that EVI1 overexpression alone confers serial replating potential on the High cell population. (I) Percentages of High population cells in primary and subsequent passage colonies derived from High populations of Gata2+/gfp::Tom and 3q21q26-EVI1::Gata2+/gfp::Tom mice. Values represent the means ± standard deviations (SDs). *P < .05; **P < .01  (unpaired Student t test). n.s., not significant.