Fig. 5.

TF mRNA expression following soluble UF exposure requires ROS production and NOX-4. HCAEC pretreated with the NOX inhibitor DPI (50μM) for 30 min followed by exposure to 0 or 50 μg/ml soluble UF were used to measure (A) extracellular H₂O₂ and (B) TF mRNA expression. (C) HCAEC were transfected with NOX-4 or scramble siRNA for 48 h. mRNA expression of the target gene NOX-4 and non-target gene GAPDH were quantified by RT-PCR. (D) HCAEC were transfected with NOX-4 or scramble siRNA for 48 h and then exposed to 0 or 50 μg/ml soluble UF for 6 h. TF mRNA expression was quantified by RT-PCR. Error bars indicate ± SE of n = 3 separate experiments with samples in duplicate. *p < 0.05, **p < 0.01, and ***p < 0.001 compared with control.