The effect of dilazep on TF mRNA expression in HUVECs stimulated with TNF, thrombin, or PMA. (A) HUVECs were stimulated with each stimulant: 1,000 U/mL TNF, 25 nmol/L thrombin, or 5 nmol/L PMA for 3 hours in the presence (+) or absence (−) of 100 μg/mL dilazep. TF mRNA expression was analyzed using the RT-PCR method as described in the Materials and Methods. The amplified cDNA was blotted onto a nitrocellulose membrane and hybridized with the [³²P]TF cDNA probe and then analyzed using an autoradiogram. (B) The data in (A) are depicted after normalization to equal amount of RNA loads based on the intensity of GAPDH mRNA. The data are expressed as the percentage of the control (each stimulant induced TF mRNA in the absence of dilazep). The values represent the mean ± SD of four independent experiments. Statistical analysis was performed using the Student's t-test (*P < .05).