Extended Data Fig. 2: PD-1 expression on colonic LTi is upregulated during activation. | Nature Metabolism

Extended Data Fig. 2: PD-1 expression on colonic LTi is upregulated during activation.

From: PD-1 signaling facilitates activation of lymphoid tissue inducer cells by restraining fatty acid oxidation

Extended Data Fig. 2

a, PD-1 expression on colonic ILCs, γδT, NKT, monocytes, and DC was examined by flow cytometry. Percentages of PD-1+ cells and their PD-1 MFI were calculated (n = 4 per group; ns, ns, ***p < 0.0001, ***p < 0.0001; ***p = 0.0008, ***p < 0.0001, ***p < 0.0001, ***p < 0.0001). b, Gating strategy for ILC subsets, including LTi, T-bet-(CCR6-) ILC3, T-bet+(NKp46-) ILC3, NKp46+ ILC3, ILC1/cNK, and ILC2. c, On day 7 of DSS-induced colitis, proportion of IL-22-producing lymphocytes in colon was analyzed (mean ± s.d.; n = 6). d, PD-1 expression between colonic LTi of B6 (Rag2+/+) and Rag2-/- mice were compared. Proportion of PD-1+ LTi and their PD-1 MFI were calculated (n = 5 per group; ***p < 0.0001, **p = 0.0027). e, IL-22 production by colonic LTi of mice in steady state and DSS-induced colitis (on day 7) was examined. Proportion of IL-22+ LTi and their IL-22 MFI were compared (n = 4 per group; **p = 0.0028, *p = 0.0189). f, On day 7 of DSS-induced colitis, forward and side scatters (FSC-A and SSC-A) of PD-1high and PD-1- colonic LTi were examined. Mean values of FSC-A and SSC-A were compared (n = 4 per group; ***p = 0.0001, ***p < 0.0001). g, On day 7 of DSS-induced colitis, IL-17A expression in PD-1high and PD-1- colonic LTi was examined. Proportion, IL-17A MFI and number of IL-17A+ LTi were compared (n = 5 per group; ***p = 0.0008, *p = 0.0454, **p = 0.0015). h, PD-1 expression on in vitro cultured LTi treated with heat-killed bacteria or PBS (left), or bacterial culture supernatant or culture medium (right) was examined. i, 7 days post C. rodentium infection, PD-1 expression on colonic LTi was examined. Proportion of PD-1+ LTi and their PD-1 MFI were calculated (n = 4 per group; ns, ns). 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, d-g, i). Ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001.

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