Extended Data Fig. 6: AFABP perturbs various signaling pathways associated with the ISC differentiation during intestinal epithelial repair.
From: Obesity impairs gut repair via AFABP-mediated iron overload in intestinal stem cells
a, Heatmap of all gene expression in the different groups. b, KEGG pathway enrichment analysis of differentially expressed genes between the OB-WT and NW-WT groups. c, Heatmap showing the expression levels of Wnt, Notch, TGF-β, and PI3K-AKT target genes in intestinal crypts from NW-WT, OB-AFABPAKO, and OB-WT groups. d, e, Relative mRNA of Wnt signaling pathways (Ctnnb1) (d) or Notch pathways (Notch1) (e). Temporal expression profiles normalized to NW-WT at DSS-0D (set as 1). Comparative analyses were conducted between NW-WT and OB-WT, as well as between Obese mice with OB- AFABPAKO and OB-WT. n = 3 independent experiments. f-i, Western blot representative images (f) and quantification of β-catenin (Wnt signaling) (g, h) and Notch intracellular domain (NICD) (i) dynamics in: NW-WT; OB-WT; OB-AFABPAKO. Temporal expression profiles normalized to NW-WT at DSS-0D (set as 1): NW-WT exhibits oscillatory β-catenin (Wnt) and Notch dynamic balance；OB-AFABPAKO restores the oscillatory pattern. n = 3 independent experiments. j, AP staining showed organoids with hAFABP addition exhibited a reduction in enterocyte differentiation and could be rescued by IWP-2 (an inhibitor of Wnt signaling), with DMSO as control. Three independent experiments were performed. k, Relative mRNA expression of indicated genes in human intestinal organoids of indicated treatment (n = 3 independent experiments). Scale bars represent 200 μm (j). Error bars indicate mean +/- SDs. P-values were calculated as indicators of statistical significance, ns signifies p > 0.05. One-way ANOVA with Dunnett multiple comparison test for (d, e, g, h, i, k). Additional data are provided in Tables S3 and S4.
