Figure 5

The effect of CAPE on the NF-κB and MAPK signaling pathway. Human MoDCs from healthy subjects were pretreated with CAPE in the indicated concentration for 2 hours, then stimulated by LPS (100 ng/mL) for 45 minutes. Cell lysate were collected and the levels of phosphorylation of (A) IκBα and (C) MAPK family (p38 MAPK, p42/44 ERK, and p46/54 JNK kinase) were assayed by Western blotting with indicated Abs. Anti-α-tubulin mAb and anti-total p38 polyclonal Ab were for internal control. (B) The pretreated MoDCs as mentioned were stimulated by LPS (100 ng/mL) for 2 hours and nuclear fractions were prepared and analyzed for NF-κB binding activity by EMSA. To assess the specificity of the binding, 100-fold excess of cold NF-κB probe or mutant probe was added to the LPS condition. One representative of at least three independent experiments is shown.