Figure 4.

The NF-κB pathway is implicated in the anti-inflammatory process of limonoid 2. (A) RAW264.7 cells were pre-treated by limonoid 2 (15, 30 μM) for 4 h and then mediated by LPS for 24 h. The expression of proteins p65, p-p65, IκBα, p-IκBα, IKKα/β, and p-IKKα/β was measured by Western blotting. (B) RAW264.7 cells were pre-treated by limonoid 2 for 4 h and then treated with LPS for 24 h. Translocation of p65 was detected by immunofluorescence, as outlined in the Methods sections. Primary anti-NF-κBp65 (1:500), secondary anti-fluorescein-coupled Goat Anti-Rabbit IgG (H + L) (1:100). * p < 0.05, ** p < 0.01, compared with the LPS group alone. The scale bar represents 10 μm.