Minimum essential medium eagle alpha modification α mem

Foetal bovine serum and Minimum Essential Medium Eagle Alpha Modification (α‐MEM) were obtained from Gibco (Sydney, Australia). Dehydrocostus lactone was purchased from Selleck (Shanghai, China), and Cell Counting Kit‐8 (CCK‐8) was obtained from Dojindo Molecular Technologies (Shanghai, China). Recombinant murine RANKL and M‐CSF were purchased from R&D Systems (Minneapolis, MN). Primary antibodies against extracellular signal‐regulated kinase (ERK), p‐ERK, c‐Jun N‐terminal kinase (JNK), p‐JNK, p38, p‐p38, p65, p‐p65, IκBα, p‐IκBα, IκB kinase β (IKKβ), p‐IKKα/β, NFATc1/NFAT2, c‐Src, and β‐actin were purchased from Cell Signaling Technology (Danvers, MA). Antibodies against cathepsin K were obtained from Abcam (Cambridge, MA). Rhodamine‐conjugated phalloidin was from Cytoskeleton (Denver, CO). Titanium particles were purchased from Alfa Aesar (#00681; Alfa Aesar, Ward Hill, MA). The TRAP staining kit, Tris, glycine, dimethylsulfoxide (DMSO), and sodium dodecyl sulfate were from Sigma‐Aldrich (St. Louis, MO) unless otherwise stated.

RAW 264.7 cells were purchased from the Korean Cell Line Bank (Seoul, Korea). Fetal bovine serum (FBS) was obtained from Atlas Biologicals (Fort Collins, CO, USA). Penicillin/streptomycin (P/S), minimum essential medium eagle alpha-modification (α-MEM), and normal serum were purchased from Gibco (Gaithersburg, MD, USA). E₂ and ALN were supplied by Sigma-Aldrich (St. Louis, MI, USA). Osteo assay surface multiple-well plates were supplied by Corning Inc. (Corning, NY, USA). Nitrocellulose membranes were purchased from Bio-Rad Laboratories, Inc. (Hercules, CA, USA). Taq polymerase was purchased from Kapa Biosystems (Woburn, MA, USA). Primary antibodies against anti-β-actin (cat no: 8432) and anti-c-Fos (cat no: sc-447) were supplied by Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Anti-NFATc1 (cat no: 556602) was supplied by BD Pharmingen, Inc. (San Diego, CA, USA), and secondary antibodies were purchased from Jackson ImmunoResearch Laboratories, Inc. (West Grove, PA, USA).

Cell culture plastics were purchased from Becton Dickinson (Lincoln Park, NJ, USA). Fetal bovine serum (FBS), phosphate-buffered saline (PBS), L-glutamine, trypsin, and antibiotics were purchased from Gibco (Life Technologies, Carlsbad, CA, USA). Trans-polydatin with a purity grade higher than 99% was provided by Glures S.r.l., Academic Spin-Off of Ca’ Foscari University of Venice (Venice, Italy). Minimum Essential Medium Eagle Alpha Modification (α-MEM) was purchased from Gibco, Thermo Fisher Scientific, Inc. (Waltham, MA, USA). All other regents used in this study were of analytical grade. Polycaprolactone was purchased from Sigma-Aldrich (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany). Chloroform and ethanol were purchased from VWR International (VWR International, Radnor, PA, USA).