Fig. 3

Expression of PID inhibits invasiveness and motility by suppressing EMT. a Cell viability of primary colon epithelium tumor cells derived from CDX2;APC;PID (#4, 7 and 26; red line) and CDX2;APC (#35, 48 and 56; black line) mice was measured by MTT assay. b Invasion assay was performed on PID⁺ (4, 7, and 26) and PID^- (35, 48, and 56) primary cells after 19 h incubation and the number of cells was counted using microscope in five random fields per chamber. c The number of migrated primary cells on both PID⁺ (4, 7, and 26) and PID^- (35, 48, and 56) was counted in five random fields per chamber under phase contrast microscope. d Real-time PCR analysis measured relative expression level of EMT genes in PID⁺ and PID^- primary cells, including Zeb1, Zeb2, Snai1, Slug, and Twist1 (Transcription factors), Fibronectin, N-Cadherin and Vimentin (Mesenchymal markers), E-Cadherin, CLDN and CRB3 (Epithelial markers) and genes related to Wnt signaling pathway (cyclin D1, cyclin D2, α-Cat, β-Cat and c-myc). e Representative immunoblot analysis of EMT markers on PID⁺ and PID^- primary cells and relative expression was quantified the intensity by Image J