Fig. 7

Smad3 transcribes THBS1 to promote the metastasis of K150 cells. A Western blot results of prominent proteins correlated with the TGF-β1 signaling pathway in bystander K150 cells at the timing (0, 15, 30, 60, 120 and 240 min) after DJ-1 stimulation. B Schematic representation of SMAD3-binding sites on THBS1 promoter. The binding site is located at -1956 ~ -1947 on the THBS1 promoter. C-D Luciferase promoter activity was measured by transfecting a PGL3 vector containing the full-length THBS1 promoter (Full) or inserts deleted either in binding site. K150 cells were induced by exogenous DJ-1 (C) or transfected with Smad3 overexpression plasmid (D). E Western blot results of TSP1 and Smad3 in K150 cells transfected with Smad3-knockdown siRNA. F-G Representative images and the statistical graph of transwell assays for bystander K150 cells transfected with siRNA targeting THBS1. Scale bars, 100 μm. H-I Representative images and the statistical graph of transwell assays for bystander K150 cells measured by LSKL. Scale bars, 100 μm. **p < 0.01, ***p < 0.001, ****p < 0.0001