Extended Data Fig. 4: Extended analysis of CD4⁺ T cells in the different gLNs of SPF and germ-free mice and activation of OT-II CD45.1 cells in all gLNs upon OVA gavage.

a–e, Frequency of TCRβ⁺CD4⁺ cells among CD45⁺ cells (a), FOXP3⁺NRP1⁺ (b) and FOXP3⁺NRP1^– cells (c) among TCRβ⁺CD4⁺ cells, RORγT⁺ cells among FOXP3⁺NRP1^– cells (d) and FOXP3^–RORγT⁺ cells among TCRβ⁺CD4⁺ cells (e) in indicated gLNs from germ-free or SPF C57BL/6 mice (n = 4). Data representative of two independent experiments. f–i, Frequency of CD25⁺ cells among CD45.1⁺ cells (f) and CD45.1⁺ cells among TCRβ⁺CD4⁺ cells (g), total CD45.1⁺ cells (h), and representative CFSE (carboxyfluorescein diacetate succinimidyl ester) dilution histogram (i) in indicated gLNs 64 h after adoptive transfer of 1 × 10⁶ naive CD45.1⁺ OT-II cells into SPF CD45.2 C57BL/6 host mice (n = 3), after gavage of OVA 48 h and 24 h before analysis.