Mouse igg i5381

Anti-RANKL antibody (ab45039; 1:1,000) was obtained from Abcam. Anti-SMAD3 (C67H9) antibody (9523, s; 1:1,000) and anti P-SMAD3 (S423/425) antibody (9520, s; 1:1,000) were purchased from Cell Signaling Technology. Monoclonal anti-HA antibody (SC-7392; 1:2,000), anti-VDR rabbit polyclonal antibody (SC-1008; 1:1,000), anti-VDR mouse monoclonal antibody (SC-13133; 1:1,000) and anti-TUBULIN antibody (SC-23948; 1:10,000) was from Santa Cruz Biotechnology. Anti-SMURF2 antibody (ABS59; 1:500) was obtained from Millipore. Monoclonal anti-FLAG antibody (F3165; 1:5,000) was from Sigma-Aldrich. Monoclonal anti-MYC antibody (AE010; 1:2,000) was from Abclonal Technology. Monoclonal anti-GFP antibody (66002-1; 1:10,000) was purchased from Proteintech. Rabbit IgG (SC-2027) was purchased from Santa Cruz Biotechnology and Mouse IgG (I5381) was from Sigma-Aldrich. Recombinant mouse TGF-β1 (catalogue no.5231) was purchased from Cell Signaling Technology; recombinant human BMP-2(120-02) was from Proteintech; β-oestradiol (oestrogen; 02194565.2) was from MP. SB525334 was purchased from Selleckchem.

Fibronectin was purified from outdated human plasma (American Red Cross, Rochester, NY) using gelatin-Sepharose (GE Healthcare, New York, NY, USA) affinity chromatography [36 (link)]. Collagen I was extracted from rat tail tendons, as described previously [35 (link)]. PDGF-BB was purchased from Thermo Fisher Scientific (Waltham, MA, USA). Antibodies and their sources are as follows (catalog or clone number in parentheses): 9D2 monoclonal antibody (mAb) [37 (link)] was a gift from Dr. Deane Mosher (University of Wisconsin, Madison, WI, USA); mouse IgG (I5381) and anti-vinculin mAb (V4505) antibodies were from Sigma-Aldrich; anti-phospho-PI3K pAb (4228) was from Cell Signaling Technology (Danvers, MA, USA); anti-β1 integrin (Ha2/5), anti-β3 integrin (2C9.G2), IgG (553961) and IgM (553957) isotype controls were from BD Biosciences (Mississauga, ON, Canada); and horseradish peroxidase (HRPO)-conjugated goat anti-mouse and goat anti-rabbit secondary antibodies were from BioRad (Hercules, CA, USA).