FIGURE 8.

NMP inhibits RANKL-mediated c-Fos expression and AP-1 activation. RAW264.7 cells were treated with RANKL alone or with NMP as indicated. a, the c-Fos protein level was analyzed by Western blotting using anti-c-Fos and anti-actin antibodies as described under “Experimental Procedures.” b, nuclear extracts from RAW264.7 cells were used to detect AP-1 activation with an ELISA-based method (TransAM AP-1) as described under “Experimental Procedures.” The specificity of the assay was monitored by using the K-562 (12-O-tetradecanoylphorbol-13-acetate-stimulated) nuclear extract and free wild-type or mutant oligonucleotides according to the manufacturer's instructions. Data are expressed as the mean ± S.D. (n = 4) from two different experiments.