Figure 5: Inhibition of Notch signalling leads to downregulation of stemness-related genes.

(a) Inhibition of the Notch pathway by addition of DBZ (1 μM; lower panel) changes the differentiation pattern and structure of the organoids, leading to distinct changes in morphology, evident by increased folding (asterisk). (b) qPCR validation of selected candidate genes that were found to be downregulated in the microarray. The stemness marker Olfactomedin4, Wnt signalling components AXIN2 and LGR6, as well as the Notch target gene HES1 were all confirmed to be downregulated in independent patient samples. Data represent mean±s.d. of three independent donors. (c) Gene Set Enrichment Analysis of the regulated genes after DBZ treatment identified by microarray, compared with a set of stem cell signature genes from mouse intestine. The correlation plot reveals a significant enrichment in the set of downregulated genes (negative t-score). (d) Venn diagram showing that 78 of the 274 ‘stem cell signature genes’ were also found to be significantly downregulated in the fallopian tube organoids on inhibition of Notch. (e) qPCR validation of selected candidate genes functionally related to cillia found to be upregulated in the microarray. ARMC4, DNAI1, FOXJ1 and LRRC were found to be consistently upregulated in four independent biological replicates. Data are presented as mean±s.d.