Extended Data Fig. 6: IL6ST signalling in IECs increases SI LP-APCs and Tregs and blunts ethanol-induced steatohepatitis.

(a–j, l–o) WT and gp130^Act/IEC littermates were fed either control (n = 3–15) or ethanol-containing (n = 3–44) Lieber DeCarli diet for 10 weeks. (f–h, l–o) A group of littermate mice was treated with tropicamide (20 mg kg⁻¹) during the last 29 days as an interventional approach (n = 6–17); 50 independent experiments. (a, b) Number of duodenal GAPs per mm². (c, d) Number of CD11c⁺ APCs per mm² in the PSI. (e) Representative IF image showing TMR-dextran (red) in GAPs, CD11c (pink), WGA (green), and UEA-I (blue) staining of duodenal GCs, and epithelial cell membranes are labelled with EpCAM (white). White drawings show GAPs in communication with CD11c⁺ APCs. Images are 5 μm sections. Scale bar=20 μm. (f–h) Isolated SI LP-immune cells were stimulated with PMA (10 ng ml⁻¹) plus ionomycin (500 ng ml⁻¹) for 4 h. Gating strategy (Supplementary Fig. 8a) (f–h). (f) Frequencies at the gate of Tregs (CD4⁺CD25⁺FOXP3⁺) within the total lymphocyte population. (g) Representative Treg scatter plots. (h) Total Treg (CD4⁺CD25⁺FOXP3⁺) number. (i) β-actin-normalized duodenal IL-10 and representative blots. (j) Cd274 mRNA amounts. (k) Volcano plot of the downregulated terms in patients with AUD from the PanglaoDB Augmented 2021 gene set. Each point represents a single term from the enrichment results. Blue text indicates pathways associated with relevant immune cells and neuronal regulation. (l, m) Representation of already described data in Fig. 4, comparing all groups in one graph. (l) Duodenal GAPs per villus. (m) Plasma ALT. (n–o) Liver Cxcl5 and Ccl2 mRNA amounts. Diagonal ticks indicate a break in the y-axis scale (f, h, i, m–o). P values were determined using two-way ANOVA with Tukey’s test (f, l–m) or BKY FDR test (h), one-way ANOVA with the BKY FDR test (j, n–o), and either the two-sided unpaired Student’s t-test (b, d), unpaired Welch’s t-test (a) or Mann-Whitney U test (c, i), or Fisher’s exact test with Benjamini–Hochberg adjustment (FDR) (k). 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.