Extended Data Fig. 8: Recovery from DSS colitis is improved by OSKM or vitamin B12.
From: Vitamin B12 is a limiting factor for induced cellular plasticity and tissue repair
(a-c) Previously published time course of bulk colonic RNAseq during DSS injury and recovery Data source: GSE13103263. Mice were treated with 2.5% DSS for 7 days as indicated. (a) CT analysis for all genes for which a score could be calculated are shown. Each boxplot represents one biologic replicate (that is, mouse), with the cages indicated as described by the authors of the original study. Dashed line represents median CT value at D0. (b) RNA-sequencing expression of the indicated genes. Values represent the scaled log2FC as compared to D0 across all samples as computed by the authors of the original study. (c) Gene set enrichment analysis of a published signature of methionine deprivation (msigdb: M13537 ref. 39) in samples from day 6 vs day 0 of DSS treatment as computed by the authors of the original study. (d-g) Recovery from DSS injury of mice in Fig. 5, with treatment as schematized in Fig. 5a; a representative subset of mice was used for analyses. (d) Periodic acid–Schiff (PAS) staining, which marks mucous produced by differentiated goblet cells, in colonic sections of mice on day 14. n = 5 mice from each group were analyzed with similar results; representative images are shown. (e) Intestinal barrier permeability as measured by FITC-dextran leak assay. Fluorescence intensity was measured in the serum of mice with the indicated treatments on day 9. n = 5 mice (control), n = 8 (DSS), n = 10 (DSS + B12). (f) Ki67 was quantified within the colonic epithelium (right) and representative immunohistochemistry images are shown (left). P-values indicate comparison to the untreated control. Scale bar is 100 µm. n = 6 mice (DSS D14), n = 5 all other groups. (g) Analysis of Lgr5 by single molecule in situ hybridization (ISH). The number of Lgr5+ clusters per 100 cells in the colonic crypts was quantified from at least 20 crypts per mouse (right) from n = 3 mice and representative images are shown (left). Asterisks indicate comparison to the untreated control. Scale bar is 50 µm. Bar graphs represent average ± SD; ****p < 0.0001 by one-way ANOVA.
