Effect of preexposure time of shear stress on TNF-α–induced increase in TF mRNA in HUVECs. (A) HUVECs were exposed to shear stress (18 dynes/cm2) or left static for 1, 3, 6, 9, 15, and 24 hours after TNF-α (100 U/mL) addition, and then lysed and processed for RT-PCR Southern blot analysis using32P-labeled TF cDNA. (B) HUVECs were exposed to shear stress (18 dynes/cm2) or left static for 3 hours before and 1, 3, and 6 hours after TNF-α (100 U/mL) addition, and then processed as in (A). (C) HUVECs were exposed to shear stress (18 dynes/cm2) or left static for 15 hours before and 1, 3, and 6 hours after TNF-α (100 U/mL) addition, and then processed as in (A). (Left) Autoradiograms of TF and GAPDH RT-PCR products; (right) densitometric analysis of TF RT-PCR products normalized with respect to corresponding GAPDH RT-PCR product levels.
