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. 2020 Mar 16;11:1406. doi: 10.1038/s41467-020-15221-z

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© The Author(s) 2020

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. 5 — a Analysis of myeloid differentiation by flow cytometry for CD11b and GR-1 in GFP+ wild-type and HMGN1-OE myeloid progenitors transduced with empty vector (EV) or AML-ETO9a-expressing retrovirus, each co-expressing GFP. n = 2 biologically independent cultures. b Serial replating assays of wild-type and HMGN1-OE cells transduced with AML-ETO9a or EV. n = 3 biologically independent replicates. c Analysis of myeloid differentiation by flow cytometry for CD11b and GR-1 in myeloid progenitors from Ts1Rhr (3 copies of Hmgn1) or Ts1Rhr_HMGN1+/− (2 copies of Hmgn1 but 3 copies of the other 30 genes in the Ts1Rhr triplication), each transduced with AML-ETO9a or EV. n = 2 biologically independent replicates. d Schematic of the experimental approach for AML-ETO9a in vivo leukemia studies. e Survival of primary recipients of wild-type or HMGN1-OE marrow transduced with AML-ETO9a or empty vector (EV). f Flow cytometry of HSPC phenotypes within GFP+ cells in spleens from moribund recipients of wild-type + AML-ETO9a or HMGN1-OE + AML-ETO9a transplants, n = 3 independent animals. g Quantification of H3K27ac levels by flow cytometry in the LT-HSC subpopulation of moribund animals’ leukemias, n = 8 independent animals for wild-type + AML-ETO9a; n = 5 independent animals for HMGN1-OE + AML-ETO9a. h LTC-IC assays showing number of colony formation units (CFU) obtained from leukemic spleens from either AML-ETO9a wild-type or HMGN1-OE transplants, n = 3 independent animals. i Quantification of GFP+ levels and cell surface markers for subpopulations within GFP+ cells in the peripheral blood 4 weeks after secondary transplantation of 100,000 GFP+ splenocytes of the indicated genotypes, n = 8 independent animals. *p < 0.0001 comparing lineage negative or CD11b+/GR-1+ populations between genotypes. j Survival after limiting dilution secondary transplantation of wild-type + AML-ETO9a or HMGN1-OE + AML-ETO9a leukemia cells at the indicated cell doses (100 k n = 8 independent animals; 10 k n = 6; 1 k n = 6, each representing two independent leukemias of each genotype). In all cases, samples compared by two-sided t test. Data are presented as mean values ± SD. Source data are provided as a Source Data file.