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. 2016 Feb 22;7:10739. doi: 10.1038/ncomms10739

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(a) Representative flow cytometry of primary animals treated with doxycycline (Dox) for 5 days and live and Lin⁻ gated. (b) Percentage of LSK population among the Lin^low compartment from primary mice (C; NHD13 and NHD13/MSI2^Dox; n=6, n=6 and n=5 from five independent experiments). (c) Percentage of the HSC (LSK⁺CD150⁺CD48⁻) compartment gated from the LSK⁺ at 1 month transplantation after 5 day dox administration in the primary animals, data representative (C; NHD13 and NHD13/MSI2^Dox same as b). (d) Chimerism (CD45.2) in the LSK compartment at 1 month transplantation after 5 day dox administration in the primary animals, data representative of two independent transplants (NHD13; n=9, NHD13/MSI2^Dox; n=8), (e) Terminal chimerism from transplants in Fig. 3 in the gated populations, (C, NHD13, NHD13/MSI2^Dox; n=11-16 combined from two independent transplants). (f) Representative flow cytometric plots from transplanted mice analysed (combined experiments from 3 and 7 months posttransplant) that were injected with Brdu 24 h and then sorted for LSK cells and gated accordingly. (g) Data represented in f, (n=3 for each group combined from two independent experiments). (h) RNA-sequencing of LSK sorted cells from primary transplanted animals (4 months posttransplantation and 3 months post MSI2 induction) before disease initiation underwent unsupervised clustering of the differentially expressed genes with human homologues, (C; n=3, NHD13; n=2, NHD13/MSI2^Dox; n=4). (i–k) Gene set enrichment analysis (GSEA) from ranked list of NHD13/MSI2^Dox/NHD13 versus Control. (l) Unsupervised clustering of the mouse NHD13 signature overlapped with MDS patients. (m) MSI2 expression (Z-score) separated based on clustering in l. (n) Survival of MDS patients based on clusters from the NHD13/MSI2 signature (l,m). Data in b–e,g and m *P<0.05, **P<0.01, ***P<0.001, are Student's t-test and the horizontal line is the mean±s.e.m and n, P values are displayed and calculated with log-rank.