Effect of FJU-C4 on LPS-induced MAPKs phosphorylation and NF-κB activation in Raw264.7 murine macrophage culture.

<p>(A) Raw264.7 macrophage cells were pretreated with FJU-C4 (10 μM) compound for 30 min and then stimulated by LPS (100 ng/ml) for 15–60 min. (B) Raw264.7 macrophage cells were pretreated with various doses of FJU-C4 compound for 30 min and then stimulated by LPS (100 ng/ml) for 15 min. The activation of ERK, p38, and JNK was analyzed by western blotting, with specific anti- p-ERK, p-p38, and p-JNK antibodies, respectively. (C) The p38 downstream targets including ATF-2 and MSK1 were also measured by using specific antibodies. (D) Raw264.7 macrophage cells were pretreated with various doses of FJU-C4 compound for 30 min and then stimulated by LPS (1 μg/ml) for 60 min. The cytoplasm and nucleus fraction were collected by a protein fraction enrichment kit. The location of NF-κB was analyzed by western blotting with specific anti-NF-κB (p65) antibody. The tubulin and lamin proteins were detected as internal control for cytoplasm and nucleus fraction, respectively.