Figure 5.
Large and small intestinal GALT possess distinct FRC landscapes. (A) Violin plots showing the expression levels of F3 and TNFSF11 by the indicated GALT FRC subclusters from ileal GALT (left plots) and large intestinal SM-ILF (right plots). (B) Confocal laser microscopy image of a large intestinal SM-ILF stained for PDPN and a TNFSF11 RNA probe. Left plot: Overview image. Right plot: Zoomed-in region of the yellow square region of the left plot. Star marks the apical side of the epithelial layer facing the intestinal lumen, yellow arrowheads mark PDPN+TNFSF11+ FRC. Representative image from six replicates from different tissue donors (with two out of six showing TNFSF11+ cells). (C) Overlay of an MRC gene signature on PP, M-ILF, and SM-ILF FRC subclusters based on the top 10 DEG expressed by ileal GALT MRC. (D–E) Bar plots showing the abundance of SED-FRC (C) and TRC (D) among GALT FRC from indicated tissues. (F) Representative confocal laser microscopy image used for the quantification of T cell zone (purple) and B cell zone (turquoise) size within a GALT follicle. 8–11 follicles from different donors were analyzed per GALT type. (G) Bar plot showing the B and T cell zone ratio per follicle. Pooled data are from eight independent experiments. (H) Nonscaled comparison of expression profiles of the indicated GALT FRC subclusters in ileal GALT and large intestinal SM-ILF. Zero (blue) indicates no overlap of the gene signatures; one (red) indicates full overlap. (I) Overlay of a curated B cell zone score on large intestinal SM-ILF FRC (left plot) and ileal GALT (right plot) FRC subclusters. (J) Overlay of a curated T cell zone score on large intestinal SM-ILF (left plot) and ileal GALT (right plot) FRC subclusters. (A, C–E, and H–J) Data are from five independently processed donors (5× PP and SM-ILF, 3× M-ILF). (C, D, and F) Statistical significance between the samples was determined using a one-way ANOVA followed by Tukey’s multiple comparison test. Dots represent individual samples, bars represent means per group, and whiskers indicate SD. ns, not significant; *P < 0.05; differences with a P value <0.05 were considered statistically significant.

Large and small intestinal GALT possess distinct FRC landscapes. (A) Violin plots showing the expression levels of F3 and TNFSF11 by the indicated GALT FRC subclusters from ileal GALT (left plots) and large intestinal SM-ILF (right plots). (B) Confocal laser microscopy image of a large intestinal SM-ILF stained for PDPN and a TNFSF11 RNA probe. Left plot: Overview image. Right plot: Zoomed-in region of the yellow square region of the left plot. Star marks the apical side of the epithelial layer facing the intestinal lumen, yellow arrowheads mark PDPN+TNFSF11+ FRC. Representative image from six replicates from different tissue donors (with two out of six showing TNFSF11+ cells). (C) Overlay of an MRC gene signature on PP, M-ILF, and SM-ILF FRC subclusters based on the top 10 DEG expressed by ileal GALT MRC. (D–E) Bar plots showing the abundance of SED-FRC (C) and TRC (D) among GALT FRC from indicated tissues. (F) Representative confocal laser microscopy image used for the quantification of T cell zone (purple) and B cell zone (turquoise) size within a GALT follicle. 8–11 follicles from different donors were analyzed per GALT type. (G) Bar plot showing the B and T cell zone ratio per follicle. Pooled data are from eight independent experiments. (H) Nonscaled comparison of expression profiles of the indicated GALT FRC subclusters in ileal GALT and large intestinal SM-ILF. Zero (blue) indicates no overlap of the gene signatures; one (red) indicates full overlap. (I) Overlay of a curated B cell zone score on large intestinal SM-ILF FRC (left plot) and ileal GALT (right plot) FRC subclusters. (J) Overlay of a curated T cell zone score on large intestinal SM-ILF (left plot) and ileal GALT (right plot) FRC subclusters. (A, C–E, and H–J) Data are from five independently processed donors (5× PP and SM-ILF, 3× M-ILF). (C, D, and F) Statistical significance between the samples was determined using a one-way ANOVA followed by Tukey’s multiple comparison test. Dots represent individual samples, bars represent means per group, and whiskers indicate SD. ns, not significant; *P < 0.05; differences with a P value <0.05 were considered statistically significant.

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