Hrp conjugated mouse or rabbit secondary antibodies

The RAW 264.7 cell line was purchased from American Type Culture Collection (ATCC® TIB-71™). C-PC was bought from Sigma-Aldrich (St. Louis, MO), dissolved in sterile water and stored at 4 °C. MTT assay kit, Alkaline phosphatase staining kit, and GAPDH antibody were also purchased from Sigma-Aldrich (St. Louis, MO). The following antibodies were bought from the company indicated in parenthesis: NFATc1 (SC-16657; Santa Cruz Biotechnology; Santa Cruz, CA), TRAP and CTSK (ab191406, ab19027; Abcam; Cambridge, United Kingdome), β3 Integrin, Caspase-3, caspase-9, c-Fos, and IκB-α (4702, 9662, 9504 S, 4384 S, 4814 S; Cell Signaling Technology; Danvers, MA), and HRP conjugated (mouse or rabbit) secondary antibodies (Santa Cruz Biotechnology; Santa Cruz, CA). Protein estimation reagents, molecular weight standards for proteins, and PAGE reagents were bought from Bio-Rad. Alizarin red solution was bought from Life-line Cell Technology (CM-0058; Fredrick, MD). Super Signal™ West Pico Chemiluminescent substrate was bought from Thermo Fisher Scientific (Waltham, Massachusetts). Rhodamine phalloidin and other chemicals were purchased from Sigma-Aldrich (St. Louis, MO).

RAW 264.7 cells were purchased from American Type Culture Collection (ATCC® TIB-71™). PG-LPS was purchased from InvivoGen (San Diego, CA, USA), dissolved in endotoxin-free water, and stored at − 20 °C. The Mem-PER™ Plus Membrane Protein Extraction Kit was purchased from Thermo Fisher Scientific (Cat. #89842; Waltham, MA, USA). Neutralizing antibodies against mouse TNF-α (AF410), recombinant mouse OPG (459MO), and mouse TNF-alpha Quantikine ELISA kit (MTA00B) were purchased from R&D Systems (Minneapolis, MN). The following antibodies were purchased from the company indicated in parentheses: RANK and TLR4 (SC-374360, SC-293072; Santa Cruz Biotechnology; Santa Cruz, CA), TNFR-1, and TNFR-2 (Abcam; Cambridge, United Kingdom). GAPDH antibody was purchased from Sigma-Aldrich (St. Louis, MO), and HRP-conjugated (mouse or rabbit) secondary antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Reagents for protein estimation, SDS-PAGE, molecular weight markers for SDS-PAGE were purchased from Bio-Rad. Chemiluminescent substrate and TLR4 inhibitor TAK-242 (508336) were obtained from Thermo Fisher Scientific (Waltham, MA).