IL-17–producing iNKT cells in pLNs are Nrp-1⁺ RTE iNKT cells. (A-B) Eight-week-old mice iNKT-enriched pLN cell suspensions were activated with PMA and ionomycin in the presence of brefeldin A and prepared for FACS analysis of IFN-γ and IL-17 production in iNKT cells. (A) Representative dot-plot analyses of IFN-γ and IL-17 expression in resting (left) and activated (middle) pLN iNKT cells, and histogram overlay (right) showing Nrp-1 expression in total (shaded gray), IFN-γ⁺ (dotted line), and IL-17⁺ (thick black line) pLN iNKT cells. (B) Nrp-1 relative MFI of total, IFN-γ⁺, and IL-17⁺ pLN iNKT cells (pooled data from 3 distinct experiments, *P < .05 by unpaired Student t test). (C-E) Absolute numbers of pLN total cells (C); total, Nrp-1⁻, and Nrp-1⁺ iNKT cells (D); and IL-17⁺ iNKT cells (E) in sham-thymectomized (sTx, light gray) and thymectomized (Tx, dark gray) mice (n = 3 for sTx and n = 4 for Tx, ns indicates not significant, *P < .05, **P < .01 by unpaired Student t test).