Figure 3

Inhibitory effect of WESS on RANKL-induced c-Fos and NFATc1 expression in BMMs. BMMs were pre-incubated with vehicle or WESS (25 μg/ml) for 3 h and further cultured in the presence of M-CSF (60 ng/ml) and RANKL (100 ng/ml) for 4 days. Total RNA and cell lysates were obtained at the indicated days. (A) c-Fos and NFATc1 mRNA levels were analyzed by QPCR. ^a P < 0.05, ^b P < 0.01, ^c P < 0.001 vs. vehicle-treated control group. (B) Total cell lysates were subjected to Western blot analysis with antibodies specific for c-Fos and NFATc1. GAPDH was used as a loading control. (C) BMMs infected with retroviruses expressing either pMX-IRES-GFP (pMX) or pMX-CA-NFATc1-IRES-GFP (pMX-CA-NFATc1) were cultured with vehicle or WESS (25 μg/ml) in the presence of M-CSF (60 ng/ml) and RANKL (100 ng/ml) for 4 days. The number of TRAP(+)MNC containing more than three nuclei with GFP expression were counted. Data represents mean ± SD of three independent experiments. ^c P < 0.001 vs. vehicle-treated control group.