Extended Data Fig. 5: IL6ST signalling in IEC regulates SI LP-APCs.
From: mAChR4 suppresses liver disease via GAP-induced antimicrobial immunity
(a–i) WT and gp130Act/IEC littermates were fed control (n = 10–11) or ethanol-containing (n = 8–12) Lieber DeCarli diet for 10 weeks. A group of littermates was treated with tropicamide (20 mg kg−1) during the last 29 days as an interventional approach (n = 5–11); 14 independent experiments. LP immune cells were isolated and stimulated with flagellin (100 ng ml−1) for 2.5 h. Gating strategy (Supplementary Fig. 6a) (a–f, h–i). (a) Frequencies at the gate of APCs (CD45+MHCII+CD11c+) within the total mononuclear phagocytes-enriched population. (b) Representative APC scatter plots. (c) Total APC (CD45+MHCII+CD11c+) number. (d) Total APC (CD45+MHCII+CD11c+) number according to CD103, CD11b, and CX3CR1 staining. (e–f) Total number of IL-23 and IL-10 expressing cells amongst all APC subsets. (g) Bacterial 16S relative to Gapdh RNA in the liver. (h–i) Total number of IL-23 and IL-10 expressing cells within all APC subsets. (j–o) MLN-immune cells from Kaede-WT and Kaede-gp130Act/IEC littermate mice fed control and alcohol-containing Lieber DeCarli diets for 10 weeks (n = 4–10) were photoconverted from green to red, and their migration to the liver was monitored; 6 independent experiments. Gating strategy (Supplementary Fig. 7a,b) (j–o). (j–m) Frequencies within total mononuclear phagocytes and numbers of the sum of the total Kaede green and red (total) (j, l) and Kaede red (MLN-migrated) (k, m) positive APCs (CD45+MHCII+CD11c+) categorized according to CD103, CD11b, and CX3CR1 staining present in the liver. (n, o) Number of total (CD3−RORγt+) ILC3 and Kaede-red MLN-migrated ILC3 (red) in the liver. Diagonal ticks indicate a break in the y-axis scale (c–i). P values were determined using two-way ANOVA with Tukey’s test (c, d–upper left panel) or BKY FDR test (d–upper right panel and lower left and right panels), or Kruskal Wallis test with the Dunn’s test (a– right panel, f) or with BKY FDR test (a-left panel, h–i), and either a two-sided unpaired Student’s t-test (g, l, o) or Mann-Whitney U test (e, j–k, m–n). Results are expressed as mean ± s.e.m. P (or q) *< 0.05, **< 0.01, ***< 0.001, ****< 0.0001. Q when applying the BKY FDR method. The illustration in o was created using BioRender.
