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. 2023 Mar 29;9(13):eadf0927. doi: 10.1126/sciadv.adf0927

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Fig. 2. — (A) Volcano plot showing differentially expressed genes from AbSC cluster. Enterocyte (blue), stem cell (green), regeneration (salmon), and fetal intestine (purple) genes are highlighted. (B) Normalized regeneration gene signature expression (42) on UMAP plot. (C) Normalized Ly6a expression on UMAP plot (top). Chromatin accessibility at Ly6a genomic locus in tdT⁺ cells isolated by FACS from Lgr5-tdT and Lgr5-Apc^f/f-tdT mice by ATAC-seq (bottom). (D) Representative images of intestinal lesions from Lgr5-Apc^f/f-tdT mice including H&E staining and tdTomato (tdT), Sca-1 (quantification on top right), Trop2, and Krt20 IHC. Scale bar, 250 μm. (E) Normalized fetal-like intestinal gene signature expression (48) on UMAP plot. (F) Average expression of differentially up-regulated (red) and down-regulated (blue) genes across fetal intestines at indicated time points of mouse development by RNA-seq (1). Heatmap representing differentially expressed genes in embryonic day 12 (E12) fetal intestines compared to adult intestines ranked by fold change; a normalized enrichment score of up-regulated genes in AbSC is shown on the right (P < 1 × 10⁻⁵). Ad, adult. (G) Normalized Tacstd2 expression on UMAP plot (top). Chromatin accessibility at Tacstd2 genomic locus in tdT⁺ cells isolated by FACS from Lgr5-tdT and Lgr5-Apc^f/f-tdT mice by ATAC-seq (bottom). (H) Pseudotime analysis of AbSCs using Monocle 3. (I) Violin plot indicated expression of fetal-like intestinal gene signature expression (48) in tdT⁺ intestinal cells from Lgr5-tdT control and Lgr5-Apc^f/f-tdT at day 10 and 28 following tamoxifen induction. P value calculated by Wilcoxon rank sum test with Bonferroni correction.