Panel A shows a schematic of the reciprocal bone marrow chimera experiment along with representative flow cytometry plots of spleen reconstitution in WT and Lingo4 knockout combinations (no explicit axes shown). Panel B shows a scatter bar plot of the number of ILC3s in the small intestine, where the x-axis represents different mouse genotypes and the y-axis represents the number of ILC3s. Panel C shows a scatter bar plot of the percentage of interleukin 22 positive ILC3s after interleukin 23 stimulation, where the x-axis represents different mouse genotypes and conditions and the y-axis represents percentage of interleukin 22 positive ILC3s. Panel D shows a scatter bar plot of the frequencies of interleukin 17 positive ILC3s following interleukin 23 stimulation across different ages, where the x-axis represents age in weeks and the y-axis represents percentage of interleukin 17 positive ILC3s. Panel E shows box plots of microbiota diversity indices including Shannon, Simpson, and Pielou for cohoused and non cohoused WT and Lingo4 knockout mice, where the x-axis represents different conditions and the y-axis represents diversity index values. Panel F shows multidimensional scaling plots based on unweighted UniFrac and Bray Curtis distances, where the axes represent MDS1 and MDS2. Panel G shows stacked bar plots of relative genus level abundance across different conditions, where the x-axis represents samples and the y-axis represents relative abundance. Panel H shows a bar plot of differentially enriched taxa identified by LEfSe analysis, where the x-axis represents taxa and the y-axis represents LDA score. Panels I, J, and K show circular phylogenetic trees representing LEfSe identified differentially enriched taxa for non cohoused WT versus Lingo4 knockout mice, cohoused versus non cohoused Lingo4 knockout mice, and cohoused versus non cohoused WT mice, respectively.
Effects of Lingo4 deficiency on intestinal lymphocytes and microbiota composition (related to Fig. 2). (A) Reciprocal BM chimeras: irradiated WT or Lingo4−/− mice were reconstituted with WT or Lingo4−/− BM, as indicated. Representative spleen reconstitution after 8 wk. n = 3; representative of two experiments. (B and C) Frequencies of ILC3s in siLP (B) and IL-22+ ILC3s (C) after IL-23 stimulation in mice shown in A. Mean ± SEM, n = 3; representative of two experiments. (D) Frequencies of IL-17+ ILC3s following IL-23 stimulation in WT and Lingo4−/− mice across ages. Mean ± SEM, n = 4–5. (E) Microbiota α-diversity of cohoused and non-cohoused WT and Lingo4−/− mice by Shannon, Simpson, and Pielou indices. n = 4. (F and G) Multidimensional scaling (MDS) plots of cohoused vs. non-cohoused WT and Lingo4−/− mice based on Unweighted UniFrac and Bray–Curtis distances (F) and relative genus-level abundance (G). n = 4. (H) Differentially enriched taxa identified by LEfSe analysis in non-cohoused T and Lingo4−/− mice represented by LDA score. n = 4. (I–K) LEfSe-identified differentially enriched taxa for non-cohoused WT vs. Lingo4−/− mice (I), cohoused vs. non-cohoused Lingo4−/− mice (J), and cohoused vs. non-cohoused WT mice (K). n = 4. (A–D) Statistical significance was assessed using one-way ANOVA with Tukey’s post hoc test. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.