HIF-1α is not required for the initiation/progression of AE9a-induced leukemia. (A) Kaplan-Meier survival curves of mice transplanted with AE9a-expressing cells or MIGR1 control (Hif-1α^Δ/Δ + AE9a, n = 7; Hif-1α^fl/fl + AE9a, n = 5; Hif-1α^Δ/Δ + MIGR1, n = 4; Hif-1α^fl/fl + MIGR1, n = 5). The log-rank (Mantel-Cox) test was used to assess statistical significance. (B) Percentage of transduced cells (GFP⁺) in PB 3 and 20 weeks after transplantation (Hif-1α^Δ/Δ, n = 17; Hif-1α^fl/fl, n = 10). (C) Percentage of myeloid cells (Gr1⁺ and/or Mac1⁺) in PB 3, 7, 10, and 20 weeks after transplantation (Hif-1α^Δ/Δ, n = 17; Hif-1α^fl/fl, n = 10). (D) Representative FACS plots of BM cells from diseased mice showing that the GFP⁺ cells are all included in the Lin^– malignant population. The differentiated populations are stained with the following antibodies: CD3 for T cells (T), B220 for B cells (B), and Gr1/Mac1 for myeloid cells (M). (E-G) Analysis of different parameters of sacrificed mice at an advanced stage of disease: myeloid cells in PB (E), Lin^– cells in PB (F) and BM (G) (Hif-1α^Δ/Δ, n = 9; Hif-1α^fl/fl, n = 5). (H) BM-cell cytospins from leukemic mice showing similar morphology of the main population in both genotypes. The scale bar = 20 μm. (I) CFU assay derived from BM cells from both genotypes (Hif-1α^Δ/Δ, n = 8; Hif-1α^fl/fl, n = 3). (J) Percentage of colonies derived from BM cells from each genotype in a competitive transplantation 1:1 (n = 4). Plots and columns represent mean ± SEM and boxes represent mean ± min-to-max values. Unless otherwise stated, 2-tailed Student t test was used to assess statistical significance. *P < .05, ***P < .001.