Intestinal region-specific Wnt signalling profiles reveal interrelation between cell identity and oncogenic pathway activity in cancer development

摘要

Model for Wnt activation. a Organoids were grown from 3 locations of the wild type (wt) murine intestine, and after transduction with stable Ctnnb1S (Cβ) a cystic morphology can be observed in all locations. b Immunoblot of CTNNB1 and loading control GAPDH in organoids grown without (wt) or with transduced Ctnnb1S (Cβ). c Number of reads matching the wild type (wt.) hotspot region of Ctnnb1 and the mutated (mut.) hotspot region of Ctnnb1:c.97T > G;109T > G;121A > G;133T > G. d Quantitative PCR of Ctnnb1 and important Wnt target genes, n = 3, *p < 0.05, **p < 0.005 (Student’s t-test), mean fold change in vertical number. All error bars represent S.E.M. e Expression of Wnt target genes [32] as average (n = 3) scaled count, centred around the location average. Right columns show summarized average logarithmic fold change (log2FC) of Cβ/wt and mean read count across locations. Indicated gene symbols are significantly differentially expressed in the average of all locations (adjusted p-value < 0.05, Wald -Test). f Gene Set Enrichment Analysis of Wnt target genes [32] in each location separately using GSEA preranked mode or (g) testing enrichment of Hallmark gene sets [50] across all locations using EGSEA