Extended Data Fig. 3: Impact of PD-1 deficiency on colonic LTi activation.

a, Percentage and number of colonic LTi in Pdcd1^+/+ and Pdcd1^-/- mice were calculated (n = 4 per group; ns, ns). b, On day 7 of DSS-induced colitis, proportion, IL-22 MFI and number of IL-22⁺ colonic LTi in Pdcd1^-/- and Pdcd1^+/- (littermate) mice were analyzed (n = 5 per group; ***p = 0.0006, *p = 0.0489, *p = 0.0425). c, On day 3 of DSS-induced colitis, proportion, IL-17A MFI and number of IL-17A⁺ colonic LTi in Pdcd1^+/+Rag2^-/- and Pdcd1^-/-Rag2^-/- mice were analyzed (n = 5 per group; ns, ns, ns). (d-h) Pdcd1^+/+Rag2^-/- and Pdcd1^-/-Rag2^-/- mice were subjected to C. rodentium infection. d, Body weight was measured daily (n = 5 per group; ***p < 0.0001). Colon length (e) (n = 5 per group; *p = 0.0436) and H&E staining indicated tissue damage (f) was assessed on day 3, g, The bacteria burdens in feces, spleen, and liver were assessed on day 3 and 7 (n = 5 per group; *p = 0.0235, *p = 0.0389; *p = 0.0242, *p = 0.0142; *p = 0.0104, *p = 0.0492). CFU, colony-forming unit. h, On day 3, proportion, IL-22 MFI and number of IL-22⁺ colonic LTi were analyzed (n = 5 per group; *p = 0.0182, **p = 0.0029, *p = 0.0331). i, Under different stimulations, proportion and IL-22 MFI of IL-22⁺ siLP LTi in Pdcd1^+/+Rag2^-/- and Pdcd1^-/-Rag2^-/- mice were examined (n = 4 per group; ***p < 0.0001, ***p < 0.0001, ***p < 0.0001; *p = 0.0267, *p = 0.0423, ***p = 0.0001). P/I, PMA+ionomycin. j, Construction strategy for Pdcd1^h/h mice. k, On day 7 of DSS-induced colitis, proportion and hPD-1 MFI of hPD-1⁺ colonic LTi in Pdcd1^h/+ and Pdcd1^h/- mice were assessed (n = 4 per group; ns, ns). l, PD-1 expression on lineage^-RORγt⁺ siLP lymphocytes of Pdcd1^fl/-Rorc-Cre and Pdcd1^fl/- (littermate) mice were compared. Data are representative of at least three independent experiments. Data are presented as the mean ± s.d., and statistical significance was determined by two-sided unpaired t-test (a-c, e, g-i, k) and two-sided paired t-test (d). Ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001.

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