Figure 5

Downregulation of TF by fenofibric acid. (A) Confluent ECV304 cells were serum starved for 24 hr and incubated with fresh serum free media containing fenofibric acid (100 μM) for 20 hr. The magnitudes of expression of TF mRNA and of β-actin were determined by Northern blot analysis as described in Methods (n = 3). A representative autoradiograph is shown on the left. Values are means ± SD (n = 6) of percent change over control without bFGF. **p < 0.01 compared to control. (B) Confluent ECV304 cells were serum starved for 24 hr and incubated with fresh serum free media containing fenofibric acid (100 μM) for 20 hr, and exposed to bFGF (10–100 ng/ml) for 4 hr. The magnitudes of expression of TF mRNA and of β-actin were determined by Northern blot analysis as described in Methods (n = 3). A representative autoradiograph is shown on the left. Values are means ± SD (n = 6) of percent change over control without bFGF. **p < 0.01 compared to control. (C) Effects of fenofibric acid (100 μM) on basal and bFGF inducible TF transcriptional activity. For determination of promoter activity raw data were tabulated as the ratio of firefly luciferase activity to that of Renilla luciferase activity (means ± SD, n = 4), and values were reported as relative to the value for the control plasmid (assigned a value of 100). FA: fenofibric acid. *p < 0.05 (D) Effects of fenofibric acid on the concentration of TF in ECV cell lysates. Confluent cells were serum starved for 24 hr, incubated with fresh serum free media with or without fenofibric acid (100 μM) for 20 hr, and exposed to bFGF (100 ng/ml) for 24 hr. Concentrations of TF in cell lysates were assayed by Western blotting. Values are means ± SD (n = 6) of fold increase over control without bFGF. FA: fenofibric acid. *p < 0.05