Effects of spermidine on NF-κB activity in LPS-stimulated BV2 microglia. (A) Cells were pretreated with the indicated concentrations of spermidine 1 h before LPS treatment for the indicated times. Total cytosolic (30 μg) or nuclear (30 μg) proteins were separated on 10% SDS-polyacrylamide gels, followed by Western blotting using anti-NF-κB p65 and IκB-α. Proteins were visualized using an ECL detection system. ERK and lamin B were used as internal controls. (B) NF-κB p65 was localized by fluorescence microscopy after immunofluorescence staining with NF-κB p65 antibody (green). Cells were stained with DAPI for visualization of nuclei (blue). Results are representative of those obtained from three independent experiments.