Effects of KMUP-1 on RANKL-induced NF-κB activation.

<p>Cells were pretreated with KMUP-1 (0–10 µM) for 24 h before the stimulation by RANKL (10 ng/ml) for 30 min. (<b>A</b>) Using Western blotting, the nuclear fractions were analyzed for protein content of p65, a subunit of NF-κB protein, and (<b>B</b>) the cytosolic fractions were analyzed for protein content of IκB-α and phosphorylated IκB-α. (<b>C, D</b>) Confocal microscopy demonstrated that KMUP-1 inhibited RANKL-induced nuclear translocation of p65 as shown by the location of anti-p65 stain within the nucleus stained with DAPI. Scale bar: 5 µm. Each value represents the mean ± S.E.M. of three independent experiments, with triplicate determinations in each experiment. ^##<i>P</i><0.01 compared with control; *<i>P</i><0.05, **<i>P</i><0.01 compared with RANKL alone.