Figure 5
TLR4/MD2 transgene expression confers LPS-responsiveness on HEK293 cells. HEK293 cells (1.5 × 105) were cotransfected with MD-2 and CD4/Toll or full-length TLR4 expression plasmids and a HBD2 promoter-linked luciferase reporter gene. Equal amounts of the corresponding empty vector were transfected into control cells such that the total amount of transfected DNA remained constant. Uniform transfection efficiencies were confirmed using a β-galactosidase reporter plasmid. 24 hours post transfection, cells were left untreated or stimulated with LPS (10 μg/ml) then lysed and reporter gene activity was quantified by luminometry. Data are expressed as HBD2 luciferase activity. Assays were performed in duplicate and are representative of at least three separate experiments. (* P < 0.05, ** P < 0.005, *** P < 0.001 vs control).