Figure 6.

Loss of FOXO results in a perturbation of the preNKP transcriptional program and the development of non-cytotoxic ILCs. (A) Principal component (PC) analysis of RNAseq data from indicated cell populations (for gating strategy see Figure 1A ) from wildtype and FOXO1,3^ΔVav mice (n = 3). The variation explained by each PC is displayed in parenthesis. (B) Hierarchically clustered heatmaps showing gene expression for selected transcription factors critical for NK/ILC development. * indicates significant differences in expression between WT and FOXO1,3^ΔVav preNKPs (FDR < 0.05, >2-fold change). (C) Volcano plot showing log2 fold change and adjusted p-value for the comparison of WT to FOXO1,3^ΔVav preNKPs. Red dots indicate genes with >2-fold change in expression and adjusted p-value < 0.05. (D) Hierarchically clustered heatmap showing row normalized expression of differential expressed genes (identified in C). Clusters I-V are indicated. (E–G) Hierarchically clustered heatmaps showing gene expression of (E) ILC1, (F) ILC2 and (G) ILC3 signature genes defined by Robinette et al. (64), * indicates significant differences in expression between WT and FOXO1,3^ΔVav preNKP (FDR < 0.05, >2-fold change). (H) Representative flow cytometry profiles showing the identification of ILC subsets in BM from WT and FOXO1,3^ΔVav mice. (I) Total number of indicated ILC subsets in BM, spleen and thymus from WT and FOXO1,3^ΔVav mice (n = 6-12). NKp46- ILC3 population had less than 50 cells per thymus in all mice from both mouse strains so was not shown. Dots represent individual analyzed animals. Bars indicate mean and SD. P-values were calculated using Mann-Whitney tests with *, ** and *** indicates p-values <0.05, <0.01 and <0.001 respectively.