Figure 6. Reduced metastases upon induction of eNK cells in Rag2^−/−Il2rg^−/− mice.

(a) Representative plots and quantification of eNK cells and MC38-GFP cells in lungs of Rag2^−/−Il2rg^−/− mice treated with control IgG or anti-asialo GM1 antibody (b) Primary tumour weight and metastatic index from WT, Rag1^−/− and Rag2^−/−Il2rg^−/− 4T1 tumour-bearing mice at day 23 after tumour cell injection. Data are representative of two pooled experiments. Each dot represents one mouse. (c) Metastatic index and India-ink-stained lungs from WT, Rag1^−/− and Rag2^−/−Il2rg^−/− 4T1 tumour-bearing mice treated with either IL-12 or Ctrl. Data are pooled from two independent studies. Each dot represents one mouse. (d) CD45⁺CD3⁻ leukocytes from lungs of 4T1 tumour-bearing WT or Rag2^−/−Il2rg^−/− mice treated with IL-12 or Ctrl were analysed for CD122⁺CD49b⁺ cells. Data represent three independent experiments (n≥4 mice per group per experiment). (e) Eomes, NKp46, NKG2D, DNAM-1, CD27, CD11b and KLRG1 expression on CD45⁺CD3⁻CD122⁺CD49b⁺ NK cells. Data represent three experiments (n≥4 mice per group per experiment). (f) Representative plot for IFN-γ production of PMA/ionomycin stimulated cells isolated from metastatic lungs of 4T1 tumour-bearing WT and Rag2^−/−Il2rg^−/− mice treated with either IL-12 or Ctrl. Frequency of NKp46⁺CD49b⁺ of total IFN-γ-producing cells. (g) Metastatic index and India-ink-stained lungs from Ifngr^−/− 4T1 tumour-bearing mice treated with IL-12 or Ctrl. Pooled data from three experiments with at least four mice per group. (h) Metastatic index and India ink-stained lungs of 4T1 tumour-bearing Rag2^−/−Il2rg^−/− mice treated with either IL-12 or Ctrl and control IgG or anti-asialo GM1 antibody. Error bars represent±s.e.m. (a,e) One-way analysis of variance and (b) unpaired Student's t-test with Welch's correction was used to determine significance (*P<0.05; **P<0.01; ***P<0.001).