Figure 5

Inhibitory effects of fHRT-BU on osteoclast formation induced by ectopic expression of NFATc1 in BMMs. (A) BMMs were infected with pMX-puro (control) or retrovirus encoding HA-tagged Ca-NFATc1 (constitutively active form) and selected with puromycin (2 μg/ml) for 24 h. BMMs infected with retrovirus were incubated with M-CSF (60 ng/ml), RANKL (150 ng/ml), and sample (10 μg/ml) for 4 days. The cells were stained for TRAP activity and the number of TRAP-positive multinuclear osteoclasts (TRAP(+)MNCs) were counted. (B) Representative microscopic pictures of the cells (100×). Data are mean ± SD (n = 3). ^a P < 0.05, versus vehicle with RANKL, ^b P < 0.05, versus HRT-BU with RANKL.