FIG 1.

Runx3 is expressed in NKC from the NKP stage, and its loss affects their maturation. (A) Western blot analysis of FACS-sorted mouse spleen NKC (NK1.1⁺ CD3⁻), IL-2-cultured mouse spleen NKC, and IL-2-cultured human blood NKC reveals expression of Runx3. (B) NKP cells express Runx3. BM of compound Runx3^{P1-AFP/+/P2-EGFP/+} mice was analyzed for GFP expression in Lin⁻ (Lin = Ter119, B220, Gr1, CD3, CD8, and CD11b) NK1.1⁻ NKG2D⁺ CD122⁺ NKPs. (C) Most NK1.1⁺ NKC express Runx3. Lin⁻ NK1.1⁺ cells of compound Runx3^{P1-AFP/+/P2-EGFP/+} mice were analyzed for GFP expression. (D) Analysis of Runx3^P1-AFP/+ and Runx3^P2-EGFP/+ expression in BM. CD122⁺ CD3⁻ BM lymphocytes were analyzed for coexpression of NKp46 and either P1- or P2-derived GFP expression. (E) Percentages of NKp46⁺ NKC in BM and spleen of WT and Runx3^−/− mice. (F) Bar graphs showing the frequency of WT and Runx3^−/− NKC subsets out of total NKp46⁺ spleen cells under resting conditions. Mean values are shown for the four maturation stages of NKC (n = 5). Significance, WT versus Runx3^−/− NKC: ∗, P < 0.05. CD11b, CD11b⁺ CD27⁻; DP, CD11b⁺ CD27⁺; CD27, CD11b⁻ CD27⁺; DN, CD11b⁻ CD27⁻.