Figure 4.

PD98059 inhibited the FGF-1-induced transactivation of the 1.2-kb matrilysin promoter in LNCaP cells. LNCaP cells (4x10⁴ cells/cm²) were transfected with a plasmid construct encoding 1.2 kb of the human matrilysin promoter driving a luciferase reporter gene. Following a 24-hour transfection, the cells were pretreated with PD98059 for 1 hour and then with FGF-1 (10 ng/ml). The cells were harvested at different time points indicated between 2 and 24 hours with 100 µl Luciferase Cell Culture Lysis Reagent from Promega. Proteins (30 µg) were assayed in 50 µl of Promega's Luciferase Assay Reagent on a luminometer. Each treatment group represents at least three wells per experiment, and at least three experiments were performed for each figure. Representative graphs are shown with error bars representing the replicates for a particular experiment.