Eighty 16-week-old C57BL/6 female mice (Animal Center of Academy of Military Medical Sciences, China) were used. Four to five mice per cage were housed under pathogen-free conditions, and were fed with standard laboratory rodent chow and tap water ad libitum. Mice were housed in plastic cages at room temperature (25°C) with a 12-h light-and-dark cycle, and acclimatized for seven days before use. All experiments were conducted in accordance with the National Institutes of Health Guide for Care and Use of Laboratory Animals and approved by the Ethics Committee of the Tianjin Medical University.
Murine receptor activator of nuclear factor Kappa-B ligand (RANKL) and murine macrophage-colony stimulating factor (M-CSF) were purchased from PeproTech (Rocky Hills, NC, USA). An immunohistochemical staining kit and 3, 3′-diaminobenzidine (DAB) substrate kit were purchased from ZSGB-BIO (Beijing, China). Dulbecco’s Modified Eagle’s Medium (DMEM), Minimum Essential Medium Alpha (MEM-α), fetal bovine serum, penicillin, streptomycin and trypsin were purchased from Invitrogen (Carlsbad, CA, USA). Other chemicals were purchased from Sigma (St. Louis, MO, USA) unless otherwise stated.
Murine receptor activator of nuclear factor Kappa-B ligand (RANKL) and murine macrophage-colony stimulating factor (M-CSF) were purchased from PeproTech (Rocky Hills, NC, USA). An immunohistochemical staining kit and 3, 3′-diaminobenzidine (DAB) substrate kit were purchased from ZSGB-BIO (Beijing, China). Dulbecco’s Modified Eagle’s Medium (DMEM), Minimum Essential Medium Alpha (MEM-α), fetal bovine serum, penicillin, streptomycin and trypsin were purchased from Invitrogen (Carlsbad, CA, USA). Other chemicals were purchased from Sigma (St. Louis, MO, USA) unless otherwise stated.