Fig. 3. AMD3100 relieves OVX-induced bone loss through reduction of osteoclast number onto the bone surface. (A) Representative micro-CT images of the distal femurs in each group are shown. (B) Each graph represents BMD, BVF, BMC, TMD, Tb.N., Tb.Sp., Cr.BMD and Cr.BMC in total analysis of cortical and trabecular bone. (C) The effects of AMD3100 on the expression of genes associated with osteoclast differentiation by quantitative real-time PCR. Total RNA was extracted from BM cultures treated with or without RANKL in the presence or absence of 25 μg/ml of AMD3100 for 3 days. Data represent mean ± SEM (Anova, Tukey’s HSD test. n = 3 per group). *P ＜ 0.05 compared with AMD3100-treated media or matched control. (D) Representative pictures showing osteoclasts in the trabecular region of OVX mice. Reduction of multi-nucleated TRAP⁺ osteoclasts was detected in BM. Arrowheads indicate active TRAP⁺ osteoclasts stained in red (original magnification, ×20, Scale bar 100 μm). (E) Histogram representing the osteoclast number/bone surface [N.Oc/BS (/mm)] and (F) the osteoblast number/bone surface [N.Ob/BS (/mm)]. Data indicate mean ± SEM (Student’s t-test. n= 4-5 per group). *P ＜ 0.05 compared with PBS-treated OVX mice.