Extended Data Fig. 7: Extended analysis of CD45.1⁺ 7B8tg cell fate in gLNs and gut upon removal of distal gLNs.

a–c, Representative flow cytometry plots of RORγT⁺ and CD45.1⁺ cell frequency among CD3⁺CD4⁺ cells in indicated gLNs of sham or ΔicLN mice as quantified in Fig. 3g (a), and frequency of Vβ14⁺CD45.1⁺ cells among CD3⁺CD4⁺ cells (b) and RORγT⁺ cells among Vβ14⁺CD45.1⁺ (c) cells in indicated gLNs of mice with sham operation (n = 12) or surgical removal of the I- and C1-gLNs (ΔicLN, n = 14) 64 h (day 3, Fig. 3f) after adoptive transfer of 4 × 10⁵ naive SFB-specific CD45.1⁺ 7B8tg cells into recently SFB-colonized SPF CD45.2 C57BL/6 Jax mice. Graph represents pooled data from three independent experiments with n = 4–5 per group each. d, e, Frequency of Vβ14⁺CD45.1⁺ cells among CD3⁺CD4⁺ cells (d) and RORγT⁺ cells among Vβ14⁺CD45.1⁺ cells (e) in indicated gLNs of Taconic sham or ΔicLN mice (n = 4) 64 h after adoptive transfer of 4 × 10⁵ naive SFB-specific CD45.1⁺ cells. Note that SFB is a stable member of the microbial community in Taconic C57BL/6 mice through parental transmission. f, g, Total numbers of CD45⁺ cells in C2-gLN of recently SFB-colonized SPF mice (f) or Taconic mice (g) at point of harvest. h–n, Frequency of Vβ14⁺CD45.1⁺ IL17a⁺ (h), Vβ14⁺CD45.1⁺RORγt⁺ (i), or Vβ14⁺CD45.1⁺ cells (j) among CD3⁺CD4⁺ cells, and of RORγT⁺ cells among Vβ14⁺CD45.1⁺ cells (k) and IL17a⁺ cells among RORγT⁺Vβ14⁺CD45.1⁺ cells (l) in lamina propria of indicated gut segments; frequency of Vβ14⁺CD45.1⁺ cells among CD3⁺CD4⁺ cells (m) and RORγT⁺ cells among Vβ14⁺CD45.1⁺ cells (n) in gLNs of sham or ΔicLN Jax mice (n = 4) 7 days after adoptive transfer of 5,000 naive CD45.1⁺ 7B8tg cells into recently SFB-colonized SPF CD45.2 C57BL/6 Jax mice. o–s, Frequency of Vβ14⁺CD45.1⁺ cells among CD3⁺CD4⁺ cells (o), RORγT⁺ cells among Vβ14⁺CD45.1⁺ cells (p) and IL17a⁺ cells among RORγT⁺Vβ14⁺CD45.1⁺ cells (q) in indicated lamina propria, or of Vβ14⁺CD45.1⁺ cells among CD3⁺CD4⁺ cells (r) and RORγT⁺ cells among Vβ14⁺CD45.1⁺ cells (s) in gLNs of sham or ΔicLN Taconic mice 7 days after adoptive transfer of 5,000 naive CD45.1⁺ 7B8tg cells (n = 4). Hash, fewer than 200 cells were recovered. *P < 0.05, **P < 0.01, ***P < 0.005 (one-tailed t-test or ANOVA); ns, not significant.