Figure 6: Rescuing effects of β-catenin expression on anti-inflammatory activity LGK974.

BEAS-2B human bronchial epithelial cells were transfected with 0.5 to 1.5 μg/well of β-catenin expression plasmid or 1.0 μg/well of control plasmid one day before pretreatment with 10 nM LGK974 for 2 hours, followed by 0.1 μg/ml LPS stimulation for 2 hours. Total cell lysates and nuclear fractions were analyzed by Western blotting for protein levels and phosphorylation. Beta-actin and TBP were used as endogenous controls for total cell lysates and nuclear fractions, respectively (A). The binding of nuclear NF-κB for its target DNA sequence, 5′-GGGACTTTCC-3′, was measured by ELISA (B). The expression of pro-inflammatory cytokine, IL-6 and IL-8, were measured by real time-qPCR (C,D). Data were compared to those of cells transfected with control plasmid and treated with LGK974 and LPS treatment (#-marked column). *p < 0.05, **p < 0.01, ***p < 0.001