Fig. 3: CS is required for cholesterol biosynthesis in colonic epithelial cells.
a The heatmap shows the relative levels of cholesterol metabolism-related genes in inflamed colonic tissues from 6-day 2.5% DSS-treated Sult2b1f/f and Sult2b1∆IEC mice. (Fold change ≥ 2, P < 0.05) (n = 4 mice/group). b Gene Set Enrichment Analysis shows the enrichment of the cholesterol synthesis gene signature in the downregulated genes (Sult2b1∆IEC vs. Sult2b1f/f mice). c Quantification of free cholesterol concentration in colonic tissues from Sult2b1f/f and Sult2b1∆IEC mice in the presence or absence of 6-day 2.5% DSS challenge using the Amplex® Red Cholesterol Assay Kit (n = 5 mice/group, one-way ANOVA with Sidak’s multiple comparison test). d Filipin III staining in HT-29 and SULT2B1-KO cells. The boxed areas are shown in the lower panels. (n = 3 fields/group, two-tailed Student’s t test). e CCK-8 assay shows the proliferation of HT-29 and SULT2B1-KO HT-29 cells in the presence or absence of cholesterol (50 μM) dissolved in β-cyclodextrin (CHOL: cholesterol; β-CYC: β-cyclodextrin). Two-way ANOVA with Tukey’s multiple comparisons test, 48 h (HT-29+β-CYC vs. SULT2B1-KO + β-CYC, *P = 0.0209; SULT2B1-KO + β-CYC vs. SULT2B1-KO + CHOL, *P = 0.0127), 72 h (HT-29+β-CYC vs. SULT2B1-KO + β-CYC, **P = 0.0012; SULT2B1-KO + β-CYC vs. SULT2B1-KO + CHOL, *P = 0.0122). f Normalized FPKM of the cholesterol biosynthesis genes in inflamed rectum tissues of 26 patients with UC and 22 healthy controls based on RNA-seq data in GSE 111889. (Two-tailed Student’s t test). g Quantification of total and free cholesterol in colonic tissues extracted by colonoscopy from healthy controls (n = 19) and patients with UC (n = 27) using the Amplex® Red Cholesterol Assay Kit. (Two-tailed Student’s t-test). h Pearson’s correlation analysis was performed between CS and total cholesterol levels, or CS and free cholesterol levels in colonic tissues from 27 patients with UC. (Two-tailed P value). 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.
