Rankl

TRIzol reagent, DNaseI enzyme, Moloney Murine Leukemia Virus reverse transcriptase, and SYBR Green positron emission tomography Master Mix were purchased from Invitrogen (Carlsbad, CA, USA). The BCA protein analysis reagent kit was provided by Pierce (Rockford, USA). Polyvinylidene fluoride was provided by Bio-Rad Laboratories (Hercules, USA). Rabbit anti-human osteoprotegerin (OPG) and receptor activator of nuclear factor-κB ligand (RANKL) were purchased from Novus Biologicals (Littleton,USA), rabbit anti-human receptor activator of nuclear factor-κB (RANK), β-Actin as well as goat anti-rabbit IgG horseradish peroxidase were provided by Cell Signaling Technology (Boston, USA). The ECL reagent kit was purchased from Amersham Pharmacia Biotech (Piscataway, USA), and all other reagents were of analytical grade.

Rapamycin (R117), Bafilomycin A1 (B1793), and 3-MA (M9281) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dulbecco’s modified Eagle’s media (DMEM): nutrient mixture F-12 (12400–024) and fetal calf serum (FCS) (10099–141) were obtained from Gibco (Grand Island, NY, USA). RIPA lysis buffer (P0013C) and protease inhibitor cocktail for general use (P1005) were acquired from Beyotime (Haimen, China). Recombinant M-CSF (400–23) and Recombinant RANKL (400–30) were obtained from PeproTech (Rocky Hill, NJ, USA). beclin1-shRNA (LV-beclin1-shRNA), beclin1 (LV-beclin1), and control (LV-control) lentiviral vector constructs were generated and produced by GenePharma (Shanghai, China). Polyclonal antibodies against beta-actin (β-actin), CD68, beclin1, LC3II/I, LC3B-II, ATG5, P62, RANKL, OPG, and Vimentin were purchased from Novus Biologicals (Littleton, CO, USA).