Figure 4
ΔTLR4 inhibits LPS-induced HBD2 expression in A549 cells. A549 cells (1.5 × 105) were transfected with pcDNA3 (empty vector) or a ΔTLR4 expression plasmid. 24 h post transfection, cells were left untreated or stimulated with LPS (10 μg/ml) for 24 h. (A) Total RNA was extracted, reverse transcribed into cDNA and used as a template in semi-quantitative PCR reactions using HBD2 and GAPDH gene-specific primers. HBD2 expression was given an arbitrary value of 1 in control cells. Data are expressed as mean +/- S.E. and are obtained from three experiments (* P < 0.05 vs control, † P < 0.05 vs control + LPS). (B) Duplicate experiments were performed in cells cotransfected with a HBD2 promoter-linked luciferase reporter plasmid. Cells were lysed and reporter gene activity was quantified by luminometry. Data are expressed as HBD2 luciferase activity (n = 3). (* P < 0.05 vs control, † P < 0.05 vs control + LPS)