Figure 3

De novo expression of F11R mRNA in inflamed endothelial cells: blockade of F11R mRNA expression in endothelial cells treated with TNFα and IFNγ by the RNA synthesis inhibitor, actinomycin. Confluent monolayers of HUVEC were maintained under Untreated conditions, or pretreated with the RNA synthesis inhibitor, actinomycin D (ActD) (5 μg/mL), in growth supplement-free media for 1 hr at 37°C. The response of HUVEC maintained in the presence of ActD alone is shown in histogram labeled ActD. The response of HUVEC treated with TNFα alone(100 u/ml) is shown in Figure 3a, and the response of HUVEC treated with IFNγ alone(200 u/ml) for 24 hrs is shown in Figure 3b. The response of HUVEC pretreated with ActD prior to 24 hr exposure to either TNFα (100 u/mL) or IFNγ (200 u/mL), is shown in the histograms labeled TNFα & ActD (see Figure 3a) or IFNγ & ActD (see Figure 3b). The F11R mRNA levels were measured by Real-Time PCR in triplicate for each condition. Values are the mean ± SEM. * P < 0.05 significant differences in F11R mRNA observed between cells exposed to TNFα or IFNγ alone vs ECs treated (or not treated) with ActD alone or ECs treated with ActD followed by their exposure to either TNFα or IFNγ.