Figure 2

We cotransfected the JEG3 cells with CRH-luciferase and B-galactosidase constructs and either empty vector (pcDNA3; 0.5 μg) or various concentrations of DN-MyD88 (Figure 2A) or DN-TRIF (Figure 2B) overnight using Fugene6. The cells were treated with cAMP (0.3 μM) or media for 5 h. Luciferase activity was determined to assess CRH promoter activation. Calorimetric β-galactosidase assay was performed to correct for the transfection efficiency. (*p < 0.01 compared to empty vector transfected cells). The data was presented as fold change in luciferase activity above media treated-empty vector transfected control wells. Each experiment was performed in triplicate or quadruplicate and repeated at least three independent times.