Figure 4
From: Efficient TGF-β/SMAD signaling in human melanoma cells associated with high c-SKI/SnoN expression
Effects of stable SKI knockdown in 1205Lu melanoma cells. A. Western analysis of SKI protein content in mock-transfected (Ø) and shSKI-transfected 1205Lu cells. B. Subconfluent melanoma cells were transfected with (CAGA)9-MLP-luc together with pRL-TK to measure transfection efficiency. Four hours post-transfection, cultures were left untreated (-) or stimulated (+) with TGF-β (10 ng/ml). Luciferase activities were measured in cell extracts 16 h post-transfection. C. Mock (Ø) and shSKI-transfected 1205Lu melanoma cells were incubated with TGF-β (10 ng/ml). RNA was extracted 8 h later. IL-11 and PTHrP expression was measured by quantitative RT-PCR. GAPDH mRNA levels served as internal controls. D. Comparative analysis of Matrigel™ invasion by mock- (Ø) and shSKI-transfected 1205Lu melanoma cells. Experiments were performed four times using duplicate samples.