Inhibitory effects of C. atrati (CA) extract on the LPS-induced pro-inflammatory cytokine expression. RAW264.7 cells were treated with the CA extract (10 μg/mL) for 30 min before LPS (10 ng/mL) treatment for 3 h. (A) The mRNA levels of IL-6, IL-1β, and β-actin were measured by RT-PCR and then quantified. (B) The cell lysates were immunoblotted with anti-IL-1β and anti-GAPDH antibodies and then quantified. (C) RAW264.7 cells were pretreated with the CA extract (10 μg/mL) for 30 min before LPS (10 ng/mL) treatment for 3 h. The cell lysates were immunoblotted with anti-phospho-IκB and anti-GAPDH antibodies. (D) RAW264.7 cells were transfected with NF-κB-luciferase reporter vector. After 24 h, cells were pretreated with the CA extract (10 μg/mL) for 30 min before LPS (10 ng/mL) treatment for 3 h, and cell lysates were analyzed for luciferase activity. (E) RAW264.7 cells were treated with the CA extract at the indicated concentrations. After 24 h, cell viability was analyzed by MTT assay. The graphs are presented as the mean ± SD of the three independent experiments. (A,B,D) * p < 0.05, ** p < 0.01 compared with indicated group. (E) n.s (p > 0.05), * p < 0.05, ** p < 0.01 compared with vehicle group.