Fig. 6. Effect of efonidipine on the nuclear translocation of NF-κB in LPS-treated BV2 cells. (A, B) Cells were co-treated with LPS (1 µg/mL) and the indicated concentrations of efonidipine for 24 h. The expression levels of cytosolic NF-κB (A) and nuclear NF-κB (B) were measured by western blotting analysis, as described in the Materials and methods. β-Actin and lamin B1 were used as loading controls for the cytosolic and nuclear fractions, respectively. Representative blots are shown. The data are presented as the mean ± SEM of at least three independent experiments (#p<0.05 vs. vehicle-treated control cells; *p<0.05 vs. LPS-treated cells). (C) Cells were co-treated with LPS (1 µg/mL) and efonidipine (10 µM) for 24 h, and immunocytochemical analysis was carried out using DAPI and anti-NF-κB p65 antibodies, as described in the Materials and methods. Representative immunofluorescence images are shown. The arrow in each image indicates the magnified cell shown in the inset. Scale bar, 50 µm. (D) Cells were co-treated with LPS (1 µg/mL) and the indicated concentrations of efonidipine for 24 h, and western blotting analysis of the cell lysates was conducted to determine the level of phospho-IκBα. β-Actin was used as an internal control. Representative blots are shown. LPS: lipopolysaccharide, Efo: Efonidipine, IκBα: inhibitory kappa Bα, NF-κB: nuclear factor-kappa B.
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