Sc 376875

Sections (5 μm in thickness) were prepared from the tissues [78 (link)]. The sections were incubated with mouse monoclonal anti-TRAP antibody (sc-376875, Santa Cruz Biotechnology, Heidelberg, Germany) in a humidified chamber O/N at 37 °C. The sections were washed with PBS and incubated with an FITC-conjugated anti-mouse Alexa Fluor-488 antibody (Molecular Probes, UK) for 1 h at 37 °C. The sections were observed and photographed using a Leica DM6 microscope (Leica Microsystems S.p.A., Milan, Italy).

Stachydrine (purity, ≥98%) and SC79 (purity, ≥97%) were purchased from Selleck Chemicals. Alpha modification of Eagle's medium (α‐MEM), penicillin/streptomycin and foetal bovine serum (FBS) were obtained from Gibco‐BRL. Recombinant mouse M‐CSF and RANKL were purchased from R&D Systems. Primary antibodies against TAK1 (#5206), phosphorylated TAK1 (p‐TAK1) (#4508), IκBα (#4814), phosphorylated IκBα (p‐IκBα) (#2859), p65 (#8242), phosphorylated p65 (p‐p65) (#3033), IκB kinase β (IKKβ) (#8943), phosphorylated IKKα/β (p‐IKKα/β) (#2697), p38 (#9212), phosphorylated p38 (p‐p38) (#4511), ERK (#4695), phosphorylated ERK (p‐ERK) (#4370), JNK (#9252), phosphorylated JNK (p‐JNK) (#4668), phosphorylated Akt (p‐Akt) (#2965), Akt (#4685), phosphorylated GSK3β (p‐GSK3β) (#9323), GSK3β (#9315), NFATc1 (#8032), PCNA (#13110) and β‐tubulin (#2146) were obtained from Cell Signaling Technology. Primary antibodies against c‐Fos (ab208942), NFATc1 (ab2796) and CTSK (ab37259) were obtained from Abcam. Primary antibody against TRAP (sc‐376875) was obtained from Santa Cruz Biotechnology and against active p65 (MAB3026) was obtained from Millipore. Cell counting kit‐8 (CCK‐8) was obtained from Dojindo Molecular Technology. TRAP staining kit, DMSO, SC‐514 and other reagents were obtained from Sigma‐Aldrich, unless otherwise indicated.