(A and C) U2OS cells were incubated with isothiocyanates and TPA for 24 h, and expression of the indicated proteins was determined by western blotting. (B) U2OS cells were plated onto a dish, scratched with a yellow pipette tip, and treated with PF573228 and TPA for 24 h. Migrating cells were photographed by phase contrast microscopy. (D–F) U2OS cells were transfected with the pGL2-MMP-9-WT, pGL2-MMP-9-mAP-1, and pGL2-MMP-9-mNF-κB reporter plasmids. The cells were then cultured with TPA and PF573228. Relative luciferase activity in the cell extract was then measured. (G) U2OS cells were transfected with negative control siRNA, siFAK1, or siFAK2, and then treated with TPA. Protein expression was determined by western blotting. (H) U2OS cells were transfected with the pGL2-MMP-9-WT reporter plasmid and siFAK2, and then treated with TPA for 24 h. Relative luciferase activity in the cell extract was then determined. (I) Cells were scratched, transfected with siFAK2, and then treated with TPA for 24 h. Migrating cells were photographed by phase contrast microscopy. Data represent the mean ± S.E. of three independent experiments. *p < 0.05 vs. control and #p < 0.05 vs. TPA.