Panel A shows genome browser–style tracks representing the analysis of ImmGen ATAC-seq data across annotated mouse lymphocyte populations, highlighting coordinated chromatin accessibility at the Rorc and Lingo4 loci (no explicit axes shown). Panel B and C display bar graphs showing the expression levels of Rorc and Lingo4, respectively, across various mouse lymphocyte populations, where the x-axis lists different lymphocyte populations and the y-axis indicates expression levels. Panels D to H present scatter bar plots illustrating Lingo4 expression in lymphocytes sorted from different tissues including small intestinal lamina propria, intraepithelial lymphocytes, mesenteric lymph nodes, spleen, and thymus, where the x-axis shows different lymphocyte subsets and the y-axis shows relative Lingo4 messenger RNA expression levels. Panel I compares Lingo4 expression in RORγt positive and RORγt negative cells sorted from small intestinal lamina propria of RorcGFP mice using a scatter bar plot. Panel J shows flow cytometry plots and a scatter bar plot depicting the gating strategy and absolute numbers of T-cell subsets in the small intestinal lamina propria of wild type and Lingo4 knockout mice. Panel K presents a scatter bar plot showing absolute numbers of T-cell subsets in the colon of wild type and Lingo4 knockout mice. Panels L to N display scatter bar plots showing the numbers of ILC3s in small intestinal lamina propria of wild type and Lingo4 knockout mice on different genetic backgrounds including Rag1 knockout, Tcrb knockout, and Tcrd knockout. Panel O shows a scatter bar plot illustrating the percentage of interferon gamma positive, tumor necrosis factor alpha positive, and granulocyte macrophage colony stimulating factor positive ILC3s from small intestinal lamina propria of wild type and Lingo4 knockout mice. Panel P presents a scatter bar plot showing interleukin 17 production by different small intestinal lamina propria ILC3 subsets. Panel Q shows flow cytometry analysis of interleukin 23 receptor expression between wild type and Lingo4 knockout ILC3s using a histogram and a scatter bar plot. Panel R displays scatter bar plots showing the frequencies of wild type and Lingo4 knockout T-cell subsets in small intestinal lamina propria eight weeks after bone marrow reconstitution of irradiated CD45.1 positive mice. Panel S presents a scatter bar plot showing Lingo4 expression in intestinal epithelial cells sorted from small intestinal lamina propria or colon and compared to small intestinal lamina propria ILC3s.
Lingo4 expression across lymphocyte populations and tissues (related to Fig. 1). (A) Analysis of ImmGen ATAC-seq data across annotated mouse lymphocyte populations showing coordinated chromatin accessibility at the Rorc and Lingo4 loci. (B and C) Expression of Rorc (B) and Lingo4 (C) across annotated mouse lymphocyte populations based on data from ImmGen consortium. (D–H) Expression of Lingo4 in lymphocytes sorted from siLP (D), IELs (E), mesenteric LNs (mLNs) (F), spleen (G), and thymus (H). Mean ± SEM, n = 3. (I)Lingo4 expression in RORγt+ and RORγt− cells sorted from siLP of RorcGFP mice. Mean ± SEM, n = 4. (J and K) Gating strategy and absolute numbers of T cell subsets in the siLP (J) and colon (K) of WT and Lingo4−/− mice. Th1, Tbet+ T cells; Th2, GATA3+ T cells; Th17, RORγt+ T cells; Treg, Foxp3+ T cells; and pTreg, RORγt+ Foxp3+ T cells. Mean ± SEM, n = 7–12; pooled from three independent experiments. (L–N) Numbers of ILC3s in siLP of WT and Lingo4−/− mice on Rag1−/− (L), Tcrb−/− (M), and Tcrd−/− (N) backgrounds. Mean ± SEM, n = 3. (O) Percentage of IFN-γ+, TNF-α+, and GM-CSF+ ILC3s from siLP of WT and Lingo4−/− mice. Mean ± SEM, n = 8; two experiments combined. (P) IL17 production by across siLP ILC3 subsets. Mean ± SEM, n = 7. Two experiments combined. (Q) Flow cytometry analysis of IL-23R expression between WT and Lingo4−/− ILC3s. Mean ± SEM, n = 5. (R) Frequencies of WT (CD45.1/2+) and Lingo4−/− (CD45.2+) T cell subsets in siLP 8 wk after bone marrow reconstitution of irradiated CD45.1+ mice. Mean ± SEM, n = 4–5; two experiments combined. (S) Expression of Lingo4 in intestinal epithelial cells sorted from siLP or colon and compared with siLP ILC3s. Mean ± SEM, n = 5. (D–S) Statistical significance was assessed using unpaired two-tailed t tests or one-way ANOVA with Tukey’s post hoc test. *P < 0.05; **P < 0.01.