Fig. 1: SULT2B1 and cholesterol sulfate (CS) levels are increased in ulcerative colitis (UC) mucosa.

a Normalized FPKM (fragments per kilobase of transcript per million mapped reads) of SULT2B1 in inflamed rectum mucosa of 26 patients with UC and 22 healthy controls based on RNA-seq data in GSE 111889. (Two-tailed Student’s t test). b SULT2B1 protein expression in the uninflamed and inflamed colonic mucosa from the same patients with UC using immunohistochemistry (n = 9, two-tailed, Wilcoxon matched-pairs signed rank test). c Quantification of CS in colonic tissues extracted by colonoscopy from healthy controls (n = 19) and patients with UC (n = 27) using LC–MS. CS concentration was calculated from the linear regression equation plotted from the standard curve and normalized with wet weight of colonic tissues. (Two-tailed Student’s t test). d Western blotting analyses were performed for SULT2B1 protein in the colon of mice challenged with the following: 3-day 2.5% DSS; 3-day 2.5% DSS + 3-day H₂O; 3-day 2.5% DSS + 6-day H₂O; 3-day 2.5% DSS + 9-day H₂O; (Con: control mice; D3: 3-day 2.5% DSS; D3 + H3: 3-day 2.5% DSS + 3-day H₂O; D3 + H6: 3-day 2.5% DSS + 6-day H₂O; D3 + H9: 3-day 2.5% DSS + 9-day H₂O) (n = 3, 2, 2, 2, 3 mice/group, respectively, one-way ANOVA with Sidak’s multiple comparison test). e Western blotting analyses were performed for SULT2B1 protein in the colon of mice challenged with the following: 5-day 0.5% DSS; 10-day 0.5% DSS; 15-day 0.5% DSS; (D5: 5-day 0.5% DSS; D10: 10-day 0.5% DSS; D15: 15-day 0.5% DSS) (n = 3 mice/group, one-way ANOVA with Sidak’s multiple comparison test). f RT-qPCR analysis of Sult2b1 mRNA in distal, media, and proximal colonic tissues with 2-, 4-, and 6-day 2.5% DSS challenge, respectively. (n = 6, 3, 3, 3 mice/group, respectively, one-way ANOVA with Sidak’s multiple comparison test). g Quantification of CS in cecal contents and colonic tissues from mice with 6-day 2.5% DSS challenge and control group using LC–MS. CS concentration was calculated from the linear regression equation plotted from the standard curve and normalized with wet weight of cecal contents or colonic tissues. (n = 6 mice/group, two-tailed Student’s t test). h Quantification of CS in colonic tissues from mice with 2-, 4-, and 6-day 2.5% DSS challenge using DESI-MSI. Cholesterol sulfate ion is at m/z 465.3044, the signal variation was normalized by the total ion current (TIC, range from m/z 400 to 500) pixel-by-pixel. The approximate lateral resolution is 200 μm (n = 5 mice/group, 2 fields/mouse, one-way ANOVA with Sidak’s multiple comparison test). Data from in vitro assays are reprehensive of at least three independent experiments. Data are shown as the mean ± SEM. Source data are provided as a Source Data file.