Effect of shear stress on stability of TF mRNA in HUVECs. (A) After treatment of HUVECs with TNF-α (100 U/mL) for 3 hours, cells were exposed to shear stress (18 dynes/cm2) or left static for 0.5, 1, 1.5, and 2 hours in the presence of actinomycin D (10 μg/mL), and then processed as in Fig 1. (B) HUVECs were exposed to shear stress (18 dynes/cm2) or left static for 3 hours before and 3 hours after TNF-α (100 U/mL) addition and further 0, 0.25, 0.5, 1, and 2 hours in the presence of actinomycin D (10 μg/mL), and then processed as in Fig 1. Data represent mean ± standard deviation (SD) of two separate experiments. (Left) The representative data from autoradiogram 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.
