Regulation of myeloproliferation and M2 macrophage programming in mice by Lyn/Hck, SHIP, and Stat5
J. Clin. Invest. Wenbin Xiao, et al. 118:924 doi:10.1172/JCI34013 [Go to this article.]

Figure 3
HSCs from dko mice exhibit increased proliferation, decreased apoptosis, and skewed differentiation into macrophages. (A) KSL CD34^– cells derived from WT and dko mice were sorted into a 96-well plate containing SCF, Flt3 ligand, and IL-11, and cultured for 4 days. *P < 0.05 versus WT values by Student’s t test. (B and C) Cell cycle analysis of KSL cells was performed by flow cytometry of cells stained with propidium iodide (B) or with Hoechst 33342 and Pyronin Y (C). Lower left, upper left, and upper right quadrants in panel C contain cells in G₀, G₁, and S/G₂/M phases, respectively. G₀, WT 55.1% ± 0.8% versus dko 34.2% ± 0.7%, P = 0.002; G₁, 31.8% ± 1.4% versus 45.0% ± 0.1%, P = 0.01; S/G₂/M, 10.1% ± 0.3% versus 20.9% ± 0.7%, P = 0.004. (D) Flow cytometric analysis indicated reduced annexin V⁺ apoptotic cells in KSL (HSC), but not in GMPs, derived from BM of dko mice. (E) Colony-forming assays showed skewed differentiation of KSL cells (100 cells seeded) of dko mice into macrophages in the presence of IL-3, IL-6, SCF, and erythropoietin. (F) Dko KSL cells (50 cells/well seeded), unlike WT cells, formed colonies in methylcellulose without growth factors, which was inhibited by anti–IL-3 and anti–GM-CSF antibodies. BFU, BFU-E; G, G-CFU; M, M-CFU; GM, GM-CFU; Mix, Mix-CFU.