Effects of BSNXD administration on CD4+ T cell-induced osteoclastogenesis. BMMs from OVX mice were co-cultured with CD4+ T cells from OVX mice for 2 days in the presence of M-CSF and then exposed to 20% OVX + mid-dose BSNXD group-derived serum or OVX + E2 group-derived serum (containing 10 nM 17β-E2) independently plus M-CSF and RANKL for 3 days. BMMs cultured with 20% OVX group serum were considered as the negative control. Subsequently, the cells were fixed and stained for TRAP. TRAP-positive multinucleated cells containing five or more nuclei were counted as osteoclasts. (A) Representative images of the TRAP-positive MNCs in each groups. (B) TRAP-positive MNCs (more than five nuclei) were counted under a microscope. Data are expressed as the mean ± standard error of the mean. **P<0.01. *P<0.05. BMMs, bone marrow-derived macrophages; OVX, ovariectomized; BSNXD, Bu-Shen-Ning-Xin Decoction; E2, estradiol; TRAP, tartrate-resistant acid phosphatase; MNCs, multinucleated cells; M-CSF, macrophage-colony-stimulating factor; CD, cluster of differentiation.