Figure 7.

FGF-2-induced cell-associated vitronectin and plasminogen system activity were abrogated by silencing the β₅ integrin with siRNA. HUVEC were transfected with β₅ siRNA for 72 hours. Samples were then treated with 50 ng/ml FGF-2 for 24 hours. (a) Samples were fixed and permeabilized with Triton X-100 to visualize total vitronectin. Samples were then labeled for vitronectin (green) and nuclei (blue). Samples were imaged in a z-stack (1 μM depth, 12 images per sample) to obtain fluorescence intensity throughout the cell. Mean intensity values were quantified with ImageJ. (b) Cell-associated vitronectin was analyzed by Western blot in cells treated with FGF-2 and genistein. Vitronectin band intensity was quantified and normalized to GAPDH and then normalized to untreated cell levels. (c) Plasmin enzymatic activity was determined using the Chromozym assay in HUVEC treated with FGF-2 and genistein. Samples were normalized to untreated cells. ^#p < 0.05 and ^∗p < 0.01 compared to those of the untreated cells. n = 3, one representative experiment of two.