Figure 5

Effects of WEL on LPS-induced NF-κB transcriptional activity via suppression of IκB-α degradation and nuclear translocation of the p65 and p50 subunits in RAW 264.7 cells. (A) Cells were transiently co-transfected with pNF-κB-Luc reporter plasmid and then were pretreated with the indicated concentrations of WEL for 12 h. LPS (1 μg/ml) was then added and cells were further incubated for 20 h. The cells were harvested and then the luciferase activities were determined by using the dual luciferase report assay system. (B, C) Cells were pretreated with the indicated concentrations of WEL for12 h and then stimulated with LPS (1 μg/ml) treatment for 30 min. Cytosol (B) and nuclear (C) protein were determined by Western blot assay using anti-IκB-α, NF-κB p50, and NF-κB p65. GAPDH and PARP1 were used as internal controls for Western blotting. A representative blot of each experiment was shown with the densitometric analysis corresponding to the mean ± SEM. of three independent experiments. n = 6 per experiment. ^*p < 0.05 vs. LPS group and ^**p < 0.01 vs. LPS group.^#p < 0. 05 vs. none LPS control group. ^##p < 0. 01 vs. none LPS control group.