Fig. 5

BBR promoted stromal expression of Wnts and ISC-mediated epithelial repair. A Diagram showing the times of DSS, BBR, and TAM treatment and colorectal sample collection. B Tracing of Lgr5⁺ cells in mouse colorectum. The samples of Lgr5-CreERT; tdTomato adult mice were sectioned and stained with DAPI. Scale bars, 50 μm. C DAB staining showed the number of Lgr5⁺ cells stained with GFP in the four mouse groups, n=3 per group. Scale bars, 50 μm. Each dot represented the average of at least five views per sample. Right panel: Quantitation results. D WB analysis of activation of β-Catenin and other signaling molecules in the colon samples of the four mouse groups. n=3 per group. E Immunostaining showed Ki67⁺ cells and activation of β-Catenin on rectal samples of normal mice, colitis mice, normal mice receiving BBR, and colitis mice receiving BBR. Scale bars, 50 μm. Each dot represented the average of at least five views per sample. n=4 per group. Right panels: Quantitation results. F Schematic diagram showing the sorting strategy of colorectal immune, epithelial, and stromal cells. G qPCR analysis of various Wnt molecules in stromal cells of the four mouse groups, n=3 per group. H Western blot analysis revealed that the colon samples of colitis mice showed a decrease in Wnt6 and Wnt10a, which was rescued by BBR. Right panels: quantitation data. n=3. Data are presented as means±SEM in (C-H). Two-way ANOVA with Fisher’s LSD post hoc analysis (α = 0.05) was applied in (C-H). p<0.05 was considered as statistically significant. Abbreviation: TAM, tamoxifen