Extended Data Fig. 10: EZH2 inhibition in PC organoids.
a, The effects of EZH2 inhibition on organoid growth. The organoids were plated as single cells and treated with or without EZH2i (10 μM GSK126) in the presence of Wnt-3A/R-spondin. The images were captured on day 7. Scale bar, 1 mm. Represntative data from three independent culture wells with similar results. b, The effect of EZH2 and porcupine double inhibition on the growth of a PASC organoid line with a moderate response to EZH2 solo inhibition (PC64). PC64 organoids were treated with EZH2i (10 μM GSK126) and/or porcupine-i (100 nM C59) from single cells. Image capture (left) and cell viability analysis with an ATP luminescent assay (right) were performed on day 7. Data are shown as mean ± s.d. Each dot represents one culture well. Scale bar, 1 mm. Data from three biological replicates. c, Gene set enrichment analysis of the first and second principal components of EZH2i-treated and untreated PC43 transcriptomes. The genes were ranked according to the loading in each principal component, and enrichment of GOBP gene sets was analysed. Top 5 gene sets with adjusted P value < 0.05 are shown. NES, normalized enrichment score. d, Gene expression changes in PC35 following EZH2 inhibition. The organoids were treated with or without 10 μM GSK126, and RNA was harvested on day 5. Differential gene analysis was performed using edgeR. Genes with fold change > 4, adjusted P value < 0.05 are shown in red, and those with fold change < 0.25, adjusted P value < 0.05 are shown in blue. e, Gene enrichment analysis of EZH2-treated PC35 organoids vs untreated organoids. The genes were ranked according to the fold change value in edgeR analysis. Top 5 GOBP terms are presented. NES, normalized enrichment score.
