Figure 4.

The developmental defect of AHR^-/- mice is cell-intrinsic. (a,b) siLP from AHR^-/- and AHR^+/+ mice were harvested and analyzed by FACS for CD4⁺ and CD8⁺ T cells (a) and dendritic cell populations (b). Cells are gated on CD3⁺ lymphocytes (a) and on CD19^-CD11c⁺ populations (b). (c) siLP cells from AHR^-/- into wildtype and wildtype into wildtype bone marrow (BM) chimeras were harvested and analyzed by FACS. Cells are gated on CD45.2⁺CD3^-CD19^-CD4^-NKp46⁺. The bottom bar represents percent of NKp46⁺NK1.1^- cells from AHR^-/- reconstituted cells, and top bar represent percent of NKp46+NK1.1- cells from AHR^+/+ reconstituted cells. Right panel represents fold increase in IL-22⁺ cells from IL-23 stimulated cells gated on CD3^-CD19^-NKp46⁺ cells from AHR^-/- and AHR^+/+ chimeras (n=4). AHR^-/- BM reconstituted mice show reduced frequencies of NKp46⁺ILC compared to WT BM reconstituted mice. The reconstitution of WT NKp46⁺ILC in irradiated WT mice was incomplete. (d) siLP cells from mixed AHR^-/- and WT bone marrow chimeras were analyzed by gating on CD45.2⁺CD3^-CD19^-CD4^-NKp46⁺ (AHR^-/-) cells and CD45.2^-CD3^-CD19^-CD4^-NKp46⁺ (AHR^+/+) cells and reconstitution of NKp46⁺ILC were compared. Bottom bar represent percent of NKp46⁺NK1.1^- cells in AHR^-/- cells, and top bar represent percent of NKp46⁺NK1.1^- cells in AHR^+/+ cells. Right panel represents fold increase in IL-22⁺ from IL-23 stimulated cells gated on CD3^-CD19^-NKp46⁺ from AHR^-/- and AHR^+/+ mixed chimeras. (n=3) (e,f) Cells isolated from the siLP of control (AHR-RORγt-cre^-/-) and AHR conditional deletion (AHR–RORγt-cre^-/Tg) mice were harvested and analyzed by FACS for NKp46⁺ ILC (e) and CD4⁺ LTi-like ILC (f) populations. Graphs show absolute numbers of cells. (n=4) Cells were also compared to absolute numbers of NKp46⁺ ILC (e) and CD4⁺ LTi-like ILC (f) populations from AHR^-/- siLP from (supplementary fig. 4)