Fig. 3

AICAr increases proportion of more mature monocytic cells in a primary sample from AML-M4 patient (normal karyotype, FLT3-ITD, NPM1mut). Bone marrow sample (Pt 07) 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 (0.1 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 and mean fluorescence intensity (MFI) of CD11b was determined by flow cytometry. Results are a representative experiment of three independent experiments. c) May-Grünwald-Giemsa stained cytospin preparations (100x magnification) from representative of two independent experiments. d) Proportion of macrophage-like cells and nucleo-cytoplasmic ratio of cells in representative experiment determined using automated image analysis of at least 500 cells per cytospin preparation (10x magnification)