Extended Data Fig. 4: IL6ST activation prevents ethanol-induced liver disease and alters GC biology.

(a–d; g–n) WT and gp130^Act/IEC littermates were fed control (n = 4–7) or ethanol-containing (n = 3–27) Lieber DeCarli diet for 10 weeks. A group of littermate mice was treated with tropicamide (20 mg kg⁻¹) during the last 29 days as an interventional approach (n = 8–19); 50 independent experiments. (a) Representative ORO-stained liver sections. Scale bar = 100 μm. (b) Quantification of ORO staining. (c) Intake of liquid diet. (d) Ethanol in plasma. (e–f) Goblet cell-specific Spdef and Retnlb mRNAs are enriched in isolated SI GCs (iGC) relative to the entire PSI; 3 independent experiments. (g) (left panel) Choline acetyltransferase (CHAT) near GCs marking cholinergic innervations, and (right panel) positive (UEA-I) GCs per mm² in the PSI. (h) Representative IF image showing CHAT expression (pink) near UEA-I (green)-labelled GCs whose membranes were stained for EpCAM (white). Nuclei were counterstained with Hoechst (blue). Yellow arrowheads indicate CHAT-expressing GCs. 40 μm sections. Scale bar=50 μm. (i) Percentage of CHAT in duodenal GAPs. (j) Representative IF image showing TMR-dextran (red) in GAPs, CHAT (pink) marking cholinergic innervations of GCs with open GAPs, Wheat Germ Agglutinin (WGA) and UEA-I staining of glycans in GCs (green and blue, respectively), epithelial cell membranes are marked by EpCAM (white). 5 μm sections. Scale bar=20 μm. (k) Expression of TUBB3 in innervated PSI GCs. (l) A representative IF image showing TUBB3-stained enteric neurons (red), GCs labelled with UEA-I (green), and epithelial cell membranes marked by EpCAM (white). Nuclei are counterstained with Hoechst (blue). 40 μm sections. Yellow arrowheads indicate cholinergic-innervated GCs. Scale bar=25 μm. (m) Number of Muc2-positive GCs per villus in the PSI. (n) Representative sections stained with Muc2 (red) showing GCs and DAPI (blue) for nuclei. 5 μm sections. Scale bar=20 μm. (o, p) Co-housed WT and gp130^Act/IEC littermates were fed ethanol-containing Lieber DeCarli diet (n = 12–13) following the chronic and binge ethanol-feeding model for 10 days. (o) Plasma ALT in co-housed mice. (p) Hepatic triglycerides. Diagonal ticks indicate a break in the y-axis scale (e, f, o, p). P values were determined using two-way ANOVA with Tukey’s test (c), one-way ANOVA with Tukey’s test (b, d, g–left panel, k and m), with the BKY FDR test (g–right panel), Krustal-Wallis with Dunn’s (k) or either a two-sided unpaired Student’s t-test (i, o and p) or Mann-Whitney U test (e, f). 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 n was created using BioRender.