Figure 5: Peripheral tissue distribution of NKT cells of DN-stage origin from E8_III-Cre⁺Rag2^fl/fl mice and NKT cells of DP-stage origin from E8_III-Cre⁺Rosa26-YFP mice.

(a) Flow cytometry of DN NKT cells of DN-stage origin (from E8_III-Cre⁺Rag2^fl/fl mice; DN origin (left)) and DN NKT cells of DP-stage origin (YFP⁺ cells from E8_III-Cre⁺Rosa26-YFP mice; DP origin (right)), in the spleen, liver, lungs, mesenteric lymph nodes (MLN), gut lamina propria (LP) and visceral adipose tissue (Adipose) (left margin). Numbers adjacent to outlined areas indicate percent DN NKT cells among viable B220^–CD4^–CD8^– lymphocytes gated as CD1d-dimer⁺TCRβ⁺ DN NKT cells. (b) Absolute number of NKT cells of DN-stage origin (left) or DP-stage origin (right) as in a. (c–e) RNA-seq analysis of genes encoding liver homing-related products in liver (c) or thymus (d,e) NKT cells of DN- or DP-stage origin (defined as in a; key), presented as FPKM values. Data are representative of at least three independent experiments per group, with three to eight mice per tissue (a,b; mean + s.e.m.), or one experiment (c–e; mean + s.e.m. of n = 3 biological replicates).