Figure 6.

Effects of crassolide on the pIRAK-1 and IkBα expression and p65 nuclear translocation in BMDC stimulated with LPS. BMDCs were treated for 2 h with either various doses of crassolide or 0.1% DMSO, followed by LPS stimulation (100 ng/mL) for 6 h. (a) Cells were harvested and total protein was extracted for Western blotting assay to detect pIRAK-1, and IkB protein levels. (b,c) GAPDH was used as a loading control, and the quantified expression levels by ImageJ software were plotted in the bar graphs. (d) NF-κB p65 DNA-binding activity in nuclear extracts of BMDC cells was determined using the TransAM kit, with the optical density at 450 nm (OD450). Data are presented as mean ± SEM of triplicate samples from one of three independent experiments. * p < 0.01, ** p < 0.01, *** p < 0.001, versus LPS + 0.1% DMSO-treated group, tested by one way ANOVA followed by Tukey’s multiple comparison test.