Figure 1

Effect of RIAA on LPS-induced inflammatory signaling pathways in RAW 264.7 cells. (A) Cells were incubated for 1 h with RIAA (0, 10 and 20 μg/ml) followed by LPS stimulation (1 μg/ml) for 2 h. Nuclear extract were analyzed for NF-κB binding by EMSA. (B) Cells were pre-incubated with RIAA (5 and 20 μg/ml) for 1 h and stimulated with LPS (1 μg/ml) for 1 h. Cell lysates were analyzed for phosphorylation of ERK1/2, p38 and JNK using western blot. (C) Cells transiently transfected with NF-κB firefly luciferase construct were incubated with RIAA (1, 5 and 10 μg/ml) or NF-κB inhibitor parthenolide (10 μM) for 1 h followed by LPS (1 μg/ml) stimulation for 8 h. The luciferase activities were determined and normalized with Renilla expression, and expressed as fold change compared to vehicle control. Data shown is representative of the experiment repeated several times. (D) Cells transiently transfected with CRE firefly luciferase construct were incubated with RIAA (1, 5 and 10 μg/ml) for 1 h followed by LPS (1 μg/ml) stimulation for 8 h. The luciferase activities were determined and corrected with Renilla expression, and expressed as fold change compared to vehicle control. Data shown is representative of the experiment repeated several times.