Skip to main content
. 2022 Dec;192(12):1712–1724. doi: 10.1016/j.ajpath.2022.08.008

Figure 2.

Figure 2

Enhancer of Zeste homolog 2 (EZH2) deletion inhibits mouse cholangiocarcinoma (CCA) cell growth and up-regulates the expression of tumor-inhibiting genes. A: Schematic illustration for establishment of EZH2-floxed CCA cells from mouse CCA tissues. B: Western blot analysis showing the levels of EZH2 and H3K27me3 in mouse #22891 and #22893 CCA cells with or without EZH2 deletion (infected by Ad-Cre or control virus [Ad-Con] for 3 days). C: The proliferation of mouse EZH2-deleted or control CCA cells as determined by WST-1 assays. D: Representative images (left) and quantification (right) of colony formation assays of mouse CCA cells with or without EZH2 deletion. E: Gene Ontology analysis of genes upregulated ≥1.7-fold (P < 0.001) in EZH2-deleted #22891 cells versus control cells. F: Heatmap illustrating the expression of the 12 identified tumor-inhibiting genes in patients with CCA from the Gene Expression Omnibus database (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE76297; accession number GSE76297). G: Quantitative RT-PCR analysis to determine the levels of the 12 tumor-inhibiting genes in EZH2-depleted and control CCA cells. H and I: EZH2-depleted human CCA cells (CCLP1) (H) and mouse #22891 CCA cells (I) were transfected with siRNAs targeting OSGIN1, PAX3, CDKN1A, or GAS1; 72 hours posttransfection, the cells were counted on an automatic cell counter after being stained with Trypan blue. ∗P < 0.05. GAPDH, glyceraldehyde-3-phosphate dehydrogenase.