Figure 2
Effects of RAS mutated expression in RAS wild-type cells and uPAR-silencing in RAS mutated cells. (A,B) Western blot analysis of uPAR and RAS expression in RAS wild-type PC9 cells transiently transfected and in SW48 cells stably transfected with RAS mutated plasmids; we used not transfected (NT) or empty vector (pRcCMV) transfected parental cell lines as controls. (C,D) H1299 and SW480 cells were transfected with uPAR or control siRNA as described in Methods. 48hr after the transfection the cells were seeded in DMEM and the adhesion and migration to VN assay was perfomed. Each bar represents the percentage of adhesion or migration to VN ± SD of three independent experiments, each performed in triplicate. uPAR knockdown was confirmed by immunoblot analysis of cell lysates from plates 48hr after the transfection. All immunoblot bands are cropped, full-length blot images are provided in Supplementary Figure S5. Bars, SDs. Asterisks indicate statistical significance, as determined by the Student t-test (***P < 0.001).
