Figure 4. Hepcidin overexpression inhibits osteoclast number and differentiate in the OVX mouse. (A, B) TRAP staining shows that overexpression Hepcidin significantly inhibits the number of osteoclasts in femoral bone in the OVX mouse; (C) Overexpression hepcidin decrease CTX concentration in the OVX mouse serum. Bone marrow macrophage, which extracted from femur and cultured with M-CSF and RANKL for 8 days, stained with TRAP to assess its differentiation. (D) TRAP staining revealed that differentiation of osteoclasts in vivo M-SCF induced in the OVX mouse; (E) The osteoclasts were counted in the overexpression hepcidin and non overexpression hepcidin OVX mice; (F) The bone pits in vivo M-SCF induced osteoclasts, which was separate from OVX mouse; (G) The bone pits in vivo M-SCF induced osteoclasts, which was separate from overexpression hepcidin OVX mouse; Quantitative polymerase chain reaction (q-PCR) analysis of the expression of bone resorption markers, including (H) Mmp9, (I) Trap, (J) Ctsk and (K) Ptk2β. Scale bar, 200 μm. The asterisks (*, **) indicate significant differences at P < 0.05, 0.01.