More genetic aberrations in human and murine GFI1-36N AML samples. (A) Percentage of patients with MDS/AML of 3 different cohorts with >2 chromosomal aberrations. Patient samples were genotyped for the presence of GFI1-36N or GFI1-36S with real time (RT)-PCR. (B) Number of patients in individual cohorts corelating to gender and age of the patients. (C) Schematic experimental setup to generate leukemic mice and the serial transplantation experiments. (D) Serial transplanted BM cells from leukemic MLL-AF9 mice and nonleukemic Lin^– cells were analyzed using RNA-seq followed by variant calling analysis. Shown is the number of variations in leukemic cells minus the number of variations in nonleukemic cells. n = 3; mean ± standard deviation. (E) Variations from (D) divided according to the functional class of mutation. Shown is the total number of mutations per genotype (left). The Venn diagram (right) represents the overlaps of missense mutations between GFI1-36S and GFI1-36N leukemic cells. (F) Scheme of the PiggyBac transposon-based mouse model. GFI1-36S or GFI1-36N mice were crossed with the PiggyBac transposon mice (Mx-Cre × Rosa26 × ATP2). Mice were injected with poly(I:C) to activate the transposon system. (G) The PiggyBac transposon-based mouse model was used to check the number of common insertion sites (CISs) of the transposon sequence. The number of CISs were calculated for each genotype. WT: n = 4, GFI1-36S (heterozygous [n = 6] and homozygous [n = 1]): n = 7, and GFI1-36N (heterozygous [n = 2] and homozygous [n = 7]): n = 9. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001. M, men; W, women; WT, wild-type.