Extended Data Fig. 1: Tγδ17 cells are stable at steady state and Vγ6⁺ Tγδ17 cells are plastic after S. typhimurium in mLN and coLP.

(a–i, k–n) Flow cytometric analysis across multiple tissues of fate-mapping mice. (a) RORγt, IL-17A, and ZsGreen expression in total γδ T cells from Rorc-Cre R26^ZSG and Il17a^Cre R26^ZSG mice. Representative of 3 experiments, n = 11. (b) Vγ6, Vγ4, and ZsGreen expression in total (left, top right) and ZsGreen⁺ (bottom right) γδ T cells from Rorc-Cre R26^ZSG mice from 2 experiments, n = 10. (c) γδ T cells (CD3⁺γδTCR⁺TCRβ⁻) from tissues of Il17a^Cre R26^TOM IFNγ-YFP mice at steady state; positive IFNγ-YFP control from coLP 4 days post-S. typhimurium (STm). Two experiments: n = 3 (FRT), 5 (siLP, coLP, lung, mLN, iLN). (d) RORγt and T-bet expression in γδ T cells from Il17a^Cre R26^ZSG mice. 2 experiments, n = 6. (e) IL-17A and IFNγ in ZsGreen⁺ γδ T cells from coLP of naïve (n = 11) and STm (n = 12) infected Il17a^Cre R26^ZSG mice; three experiments. (f) IL-17A⁻IFNγ⁺ ZsGreen⁻ γδ T cells from coLP. n = 9 naïve, 10 STm; two experiments. (g) Frequency of IL-17A⁺IFNγ⁻, IL-17A⁺IFNγ⁺, IL-17A⁻IFNγ⁺ Vγ4⁺ ZsGreen⁺ γδ T cells from coLP. n = 9 naïve, 10 STm; two experiments. (h) Cytokine production by ZsGreen⁺ γδ T cells from mLN. Three experiments, n > 10. (i) Vγ4 or Vγ6 versus IFNγ-YFP in TOM⁺ γδ T cells from coLP of naïve and STm infected mice from 3 experiments, n = 10. (j) Splenic CFUs post-aroA⁻ STm infection. n = 4/timepoint; one experiment. (k) IFNγ⁺ Vγ6⁺ ZsGreen⁺ γδ T cells after actA⁻ L. monocytogenes (LmOva actA-) infection. n = 2-4/timepoint; one experiment. (l) IL-17A and IFNγ in CD4⁺ ZsGreen⁺ and Vγ6⁺ γδ T cells 15 days post-C. rodentium. Two experiments, summary from one, naïve (n = 4), Cr (n = 6). (m) Normalized RORγt in Vγ6⁺ ZsGreen⁺ γδ T cells from naïve (n = 11) and STm (n = 15) infected mice; three experiments. (n) IL-22 in CD4⁺ ZsGreen⁺ and Vγ6⁺ γδ T cells post-aroA⁻ STm infection. n = 4/timepoint; one experiment. Cytokine expression in (a, e–h, k, l, n) measured after 4 h PMA/ionomycin stimulation. All results represent mean ± s.e.m. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns, not significant (two-tailed unpaired Student’s t-test). Numbers in flow plots represent percentages of cells in the gate.