Effect of EPA on NF-κB p65, iNOS and cytokines protein levels of in RAW264.7 cells. (A) Effect of EPA on the protein expression level of NF-κB p65 and IκB-α in RAW264.7 cells. The expression of protein was analyzed by Western blot. The β-actin was used as an equal loading control; (B) Immunofluorescence staining demonstrating the effects of DMSO (equivalent volume 3.0 μmol EPA, as control) or EPA (3.0 μmol) on subcellular localization of NF-κB p65 in RAW264.7 cells. Visualization of the NF-κB p65 fluorescence was recorded by using a LSCM, bar = 50 µm (×400); (C) EPA regulated the protein expression of iNOS; (D) EPA regulated cytokines protein levels of RAW264.7 cells. Cells were treated with DMSO (equivalent volume 3.0 μmol EPA, as control) or EPA at the various concentrations (1.2, 1.8, 2.4 and 3.0 μmol) for 24 h in the presence or absence of pretreated 10 μM PDTC. (Means ± SD, * p < 0.05 and ** p < 0.01 compared with the control). Each bar is representative of 6 independent experiments, and data were analyzed by ANOVA and Duncan’s multiple range tests.