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Figure 5: Dkk-3 inhibits epithelial-mesenchymal transitions and transcriptional factors, decreasing tumorigenesis and cellular motility/migration. (a) EMT-related marker, E-cadherin (epithelial marker) expression for Dkk-3-transfected SaOS-2 OS cells compared to control vector was determined by real-time RT-PCR. Columns, mean from 3 independent experiments; bars, SE. Western blot analysis showing expression of E-cadherin along with other epithelial markers, including Keratin 8, Keratin 18 for both subsets. Beta actin utilized as housekeeping gene. (b) EMT-related markers, N-cadherin (mesenchymal marker) fold changes in mRNA level comparing SaOS-2 OS control to Dkk-3-transfected SaOS-2 OS cells. Columns, mean from 3 independent experiments; bars, SE. Western blot analysis of mesenchymal markers (N-cadherin, fibronectin) for both subsets. (c) Immunofluorescent microscopy of N-cadherin and E-cadherin staining in transfected SaOS-2 OS cells; magnification 400x. (d) Real-time RT-PCR of transcription factors (Snail, Twist, and Slug) in transfected SaOS-2 OS cells; columns, mean from 3 independent experiments; bars, SE. Western blot analysis of Snail, Slug, and Twist in transfected SaOS-2 OS cells.