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Fig. 1 | Journal of Hematology & Oncology

Fig. 1

From: A four-stage model for murine natural killer cell development in vivo

Fig. 1

Four in vivo developmental stages of murine NK cells in the bone marrow. A NKp46 levels in the four populations based on NK1.1 and CD49b within LinCD122+ cells in the BM. B NK1.1 and CD49b expression in LinCD122+NKp46 or LinCD122+NKp46+ cells in the BM. C Gating strategy that reveals four distinguishable subpopulations of LinCD122+ cells (I–IV) based on the NK1.1, CD49b and NKp46 markers. D CD11b and CD27 expression in populations I–IV. E Characteristics of four developmental stages of murine NK cells. Single-cell suspension from BM cells was prepared from wild-type mice and stained with indicated cell markers for flow cytometry. For examining IFN-γ and TNF-α, cells were stimulated with a leukocyte activation cocktail containing GolgiPlug for 4 h. The expression levels of the markers are scaled to: – (No/low expression, expression levels < 5%), + (intermediate, expression levels = 5%–50%), and +  + high, expression levels > 50%). F, H NK cell populations I–IV were sorted from NKp46+/GFP reporter mice and 5 × 104 cells were seeded into a 96-well plate and cultured in the presence of IL-15 (50 ng/ml) for 14 days. The cells were then harvested and analyzed using flow cytometry. Summary data (F) and representative dot plots (H) are shown for NK cell populations I–IV from NKp46+/GFP reporter mice after two weeks of ex vivo differentiation (n = 3 per group). G, I NK cell populations I–IV from CD45.1 mice were sorted and 1 × 104 to 1 × 105 cells were injected intravenously into Rag2−/−Il2rg−/− mice. The presence of transferred cells was analyzed eight weeks after adoptive transfer. Summary data (G) and representative dot plots (I) are shown for NK cell populations I–IV in the BM eight weeks after adoptive transfer (n = 3 per group)

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