Figure 5

Inhibition of LPS-induced nuclear accumulation of NF-κB p65 and degradation of IκB-α by EEPC pre-treatment in RAW 264.7 macrophages. Cells were treated with 8 μg/ml EEPC for 1 h before LPS treatment (0.5 μg/ml) for the indicated times. (A) Nuclear and cytosolic proteins were subjected to 10% SDS-polyacrylamide gels followed by Western blotting using anti-NF-κB p65 and anti-IκB-α antibodies. Nucleolin and actin were used as internal controls for the nuclear and cytosolic fractions, respectively. (B) Cells were pre-incubated with EEPC 1 h before stimulation with LPS for 30 min. Nuclear extracts were then assayed for NF-κB activity by EMSA (N.S.; non specific).