Fig. 2

AICAr induces differentiation and accumulation of macrophage-like cells in a primary sample from AML-M2 patient (normal karyotype, FLT3wt NPM1wt, primary refractory). Bone marrow sample (Pt 14) was seeded at concentration 0.4 × 10⁶/mL in medium supplemented with 50 ng/mL IL-3, IL-6, SCF and FLT3L and incubated with AICAr (0.2 and 0.4 mM), ATRA (1 μM), combination of AICAr (0.2 mM) and ATRA (1 μM), or brequinar (500 nM) for 72 h. a) Flow cytometric analysis of CD11b⁺CD34, CD11b⁺CD45⁺, CD45^highCD34⁻ and CD64⁺ populations. Percentage of cells in population of interest is indicated in respective gates. b) The number of viable cells was determined by trypan blue exclusion. Mean fluorescence intensity (MFI) of CD11b and percentage of CD64 and CD11b positive cells were determined by flow cytometry. c) May-Grünwald-Giemsa stained cytospin preparations (100x magnification). d) Proportion of macrophage-like cells in samples and nucleo-cytoplasmic ratio of cells determined using automated image analysis of at least 1000 cells per representative cytospin preparation (10x magnification). e) Cell cycle analysis of propidium-labeled cells. Results are representatives of three independent experiments