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 5
Stat5 activation is required for the increased proliferation and macrophage differentiation of SHIP^–/– KSL cells. (A) KSL CD34^– cells derived from WT and SHIP^–/– mice were sorted into a 96-well plate containing SCF, Flt3 ligand, and IL-11 and cultured for 3 days. *P < 0.05 between WT and mutant. (B) Colony-forming assays showed skewed differentiation of KSL cells of SHIP^–/– mice into macrophages in the presence of IL-3, IL-6, SCF, and erythropoietin (left panel). SHIP^–/– KSL cells formed colonies in the absence of growth factors (right panel). (C–E) Stat5 in SHIP^–/– c-Kit⁺Lin^– cells (C) and F4/80⁺ lung macrophages (C–E) was constitutively activated, as shown by flow cytometry (C), immunohistochemistry (D), and confocal microscopy (E) using anti–p-Stat5 (Tyr694). Original magnification, ×400 (D and E). (F and G) SHIP^–/– and WT KSL cells were transduced with a bicistronic retroviral vector encoding DN Stat5, DN Stat3, or empty vector, together with GFP, and cultured in SCF, Flt3 ligand, and IL-11. Ratios of cell numbers normalized against those of empty vector–transduced cells are plotted. ^#P < 0.05 compared with cells transduced with empty vector. (F). Transduced cells (150 cells seeded) were cultured to form colonies in methylcellulose medium containing IL-3, IL-6, SCF, and erythropoietin (G).