TGFβ signaling enhances enrichment of RORγt⁺ iNKT cells in immune and peripheral tissues. Flow cytometry analysis of the presence of iNKT cell subsets in the (A) thymus and (B) pLNs of CD4-Cre; Stop^flox/floxTgfbRI^CA (TGFβRI^CA) mice and WT littermate control 6- to 8-week-old mice. Invariant NKT cells were identified by excluding B220⁺ and CD8⁺ cells and gating on CD1d tetramer⁺ TCRβ⁺ cells. (C) Mean absolute numbers ± SEM of RORγt⁺ iNKT cells from the thymus and pLNs. Representative plots and combined data from 7 experiments totaling 10 to 12 mice in each group. P values were calculated using unpaired Student t test (*P < .05; **P < .01; NS indicates not significant). (D) Percentage ± SEM of RORγt⁺ iNKT cells among all iNKT cells in the spleen, mesenteric lymph nodes (mLNs), liver, lung, blood, and skin of TGFβRI^CA mice and WT littermate control mice. P values were calculated using unpaired Student t test (*P < .05; **P < .01; ***P < .001). (E) Flow cytometry analysis of CCR7 on RORγt⁺ and conventional RORγt⁻ iNKT cells from the pLNs. Graph illustates the mean of fluorescence intensity (MFI) of CCR7 staining on RORγt⁺ iNKT cells ± SEM. Representative plots and combined data from at least 6 experiments totaling 10 to 12 mice in each group (A-B). Representative plots and combined data from 3 to 6 experiments totaling 4 to 8 mice in each group (D-E).