Fig. 5: Loss of NR2F6 reduces the splenic cDC1 and macrophage compartment.

A Representative dot-plots of splenic DC (CD11c⁺MHC-II⁺) populations (out of CD45⁺Lin^-BB20^-Ly6C^-F4/80^-) in wild-type (Nr2f6^+/+) or Nr2f6-deficient (Nr2f6^−/−) mice. B Representative dot-plots of splenic XCR1⁺cDC1 (XCR1⁺CD11b^-) populations derived from CD45⁺Lin^-BB20^-Ly6C^-F4/80^- CD11c⁺MHC-II⁺ cells in wild-type (Nr2f6^+/+) or Nr2f6-deficient (Nr2f6^−/−) mice. C Quantification of the frequencies of parent and of CD45⁺ splenic XCR1⁺cDC1 cells in wild-type (Nr2f6^+/+) or Nr2f6-deficient (Nr2f6^−/−) mice. D Quantification of IL-15Rα MFI in splenic DC populations in wild-type (Nr2f6^+/+) or Nr2f6-deficient (Nr2f6^−/−) mice. E Representative dot-plots of splenic Ly6C^-B220^-, monocyte (Ly6C⁺B220^-), plasmacytoid DC (Ly6C⁺B220⁺), and B cell (Ly6C^-B220⁺) populations in wild-type (Nr2f6^+/+) or Nr2f6-deficient (Nr2f6^−/−) mice. F Representative dot-plots of splenic macrophage (CD11b^midF4/80⁺) populations derived from Ly6C^-B220^- in wild-type (Nr2f6^+/+) or Nr2f6-deficient (Nr2f6^−/−) mice. G Quantification of frequency of parent and of CD45⁺ total splenic macrophages (CD45⁺Lin^-B220^-Ly6C^-CD11b^midF4/80⁺) in wild-type (Nr2f6^+/+) or Nr2f6-deficient (Nr2f6^−/−) mice. H Quantification of IL-15Rα expression (MFI) in splenic macrophage populations of wild-type (Nr2f6^+/+) or Nr2f6-deficient (Nr2f6^−/−) mice. I Schematic overview of the experimental setup, splenic NK cells of wild-type (Nr2f6^+/+) or Nr2f6-deficient (Nr2f6^−/−) mice were isolated and expanded in vitro for 7 days with 50 ng/ml IL-15. NK cells were left unstimulated (M) or were stimulated for 5 hours with IL-12 + IL-18, α-NKp46, or co-cultured with B16-F10 tumor cells, IFNγ and TNFα cytokine levels were measured. J Frequency of IFNγ producing NK cells, K MFI of IFNγ expressing NK cells and L frequency of TNFα producing NK cells were quantified for wild-type (Nr2f6^+/+) or Nr2f6-deficient (Nr2f6^−/−) NK cells. A–H Representative data are shown as pooled experiments of two independent experiments n = 8. J–L The representative data shown are from one independent experiment out of two replicative experiments, with n = 4 per group and experiment. Each dot represents the data of an individual mouse. Results are shown as mean ± SD. The normality of data was evaluated by the Shapiro–Wilk test. An asterisk indicates statistically significant differences between genotypes calculated using Student’s t-test, or Mann–Whitney U test. A p value < 0.05 was considered statistically significant. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.