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. 2021 Apr 30;12:2482. doi: 10.1038/s41467-021-22750-8

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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. 3 — a Kaplan–Meier survival curve of irradiated CD45.1⁺ wild-type mice transplanted with 2 × 10⁵ Cux1^+/−;Flt3^ITD (n = 7) or Flt3^ITD (n = 5) bone marrow cells, along with the same number of CD45.1⁺ wild-type support bone marrow cells. Bone marrow cells from two Cux1^+/−;Flt3^ITD mice diagnosed with AML at 30 weeks of age were injected into three or four irradiated recipients. Flt3^ITD donor cells were obtained from two animals from a similar age as Cux1^+/−;Flt3^ITD mice and injected into two or three recipients. Median survival in Cux1^+/−;Flt3^ITD cell recipients was 85 days. P = 0.0016, log-rank test. b Representative photograph of an enlarged spleen (left) and pale femur (right) from AML-diseased 85-day-old mouse transplanted with Cux1^+/−;Flt3^ITD bone marrow cells. c Blood count parameters from mice transplanted with cells of the indicated genotypes at 12 weeks post transplantation. Higher white cell counts due to increased monocyte and neutrophil numbers are seen in mice transplanted with Cux1^+/−;Flt3^ITD bone marrow cells. d–g MGG-stained blood smears (d) showing abnormal circulating neutrophils and monocytes and H&E-stained bone marrow (e), spleen (f) and liver (g) sections showing myelomonocytic cell tissue infiltration compromising normal tissue architecture. Five mice were assessed. h MGG-stained bone marrow cytospin showing AML blasts from an 85-day-old mouse transplanted with Cux1^+/−;Flt3^ITD bone marrow cells. Scale bars; black = 20 µm, red = 50 µm, white = 100 µm. Five mice were assessed. i Flow cytometric plots showing increased proportion of myeloid (Gr1⁺Mac1⁺, red) cells within blood, bone marrow and spleen. The proportions of B220⁺ B cells (black) and CD4⁺CD8⁺ T cells (blue) are also shown. j Agarose gel image showing PCR genotyping for Flt3^ITD (top) and Cux1 (bottom) alleles in Flt3^ITD (n = 8, lanes 1-8) and Cux1^+/−;Flt3^ITD (n = 8, lanes 9-16) mice. M, DNA ladder; neg, no DNA template. Identity of PCR products is shown on the left. Each circle represents one mouse. Plots show mean + s.e.m. Two-tailed, unpaired Mann–Whitney test.