Extended Data Fig. 7: Top genes that correlate with CHRM4 expression.
From: mAChR4 suppresses liver disease via GAP-induced antimicrobial immunity
(a–b) RNA-seq analysis was performed as in Fig. 1 (AUD, n = 44; controls, n = 16). (a) Heatmaps displaying Pearson correlation (r) values for the top 30 genes that positively and negatively correlate with CHRM4 mRNA. (b) Linear regression and correlation coefficients (R) depicting co-expressed gene pairs across the samples. The straight, bold line represents the linear regression fit, and the lines above and below the bold line represent the 99% prediction bands. (c–d) Chrm4 integrated density in GCs determined by RNAscope from 8- to 12-week-old Chrm4ΔGC and WT littermates (n = 3; 2 independent experiments). (d) Representative image showing Chrm4 mRNA (red), EpCAM-stained epithelial cell membranes (white), and DAPI-stained nuclei (blue). Yellow arrowheads: Chrm4 expressing GCs. Scale bars=20 μm. (e–j) gp130Act/IEC and Chrm4ΔGCgp130Act/IEC littermates were fed ethanol-containing Lieber DeCarli diet (n = 10–14) following the chronic and binge ethanol-feeding protocol for 10 days, 11 independent experiments. (e) Hepatic Acta2 mRNA. (f) Plasma ethanol. (g–h) Hepatic Adh1 and Cyp2e1 mRNAs. (i–j) Duodenal Muc2 and Spdef mRNAs. Diagonal ticks indicate a break in the y-axis scale (e). P values from two-tailed t-tests on regression coefficients from multiple (a) or simple linear regression (b) and by either a two-sided unpaired Student’s t-test (f, h–i) or unpaired Welch’s test (c–e) or Mann-Whitney U test (g, j). Results are mean ± s.e.m. *P < 0.05, ****P < 0.0001. The illustrations in b and j were created using BioRender.
