Fig. 7.
Antioxidants block NF-κB induction in FLS treated with Ti particles. FLS were treated with control media or media containing Ti particles (5 × 106 particles/ml) in the presence or absence of 100 μM antioxidant butylated hydroxyanisole (BHA) for various times. A: 15 min before assaying, the cultures were washed with PBS and incubated with 2′,7′-dichlorofluorescin diacetate (5 μg/ml). Relative fluorescence units were detected using a microplate fluorometer after washing to remove free dye. B: total cell protein extracts were harvested at various times for Western blot for IκB. β-Actin was used as a loading control. C: nuclear protein extracts were obtained for use in gel mobility shift assays using an NF-κB consensus binding sequence.