Tartrate resistant acid phosphatase staining solution

Nardosinone (Figure 1A), purchased from Must Biotechnology (Chengdu, China), was dissolved in dimethyl sulfoxide (DMSO). Alpha modification of Eagle’s minimum essential medium (α-MEM), fetal bovine serum (FBS), and penicillin/streptomycin were purchased from Gibco BRL (Gaithersburg, MD, United States). Recombinant murine M-CSF and RANKL were purchased from R&D Systems (Minneapolis, MN, United States). Tartrate-resistant acid phosphatase (TRAP) staining solution, Triton X-100, and 4′,6-diamidine-2′-phenylindole dihydrochloride (DAPI) were obtained from Sigma-Aldrich (St. Louis, MO, United States). FITC phalloidin was obtained from Yeasen biotech Co., Ltd (Chengdu, China). Primary antibodies targeting GAPDH, IκBα, phospho-Akt, Akt, phospho-ERK1/2, ERK1/2, phospho-JNK1/2, JNK1/2, phospho-p38, p38, phosphor-PLCγ2, PLCγ2, c-Fos and NFATc1 were purchased from Cell Signaling Technology (Danvers, MA, United States). Dichlorofluorescin diacetate (DCFDA) cellular ROS detection assay kits were obtained from Beyotime Institute of Biotechnology (Jiangsu, China). LPS from P. gingivalis was purchased from InvivoGen (San Diego, CA, United States).

Alpha minimum essential medium (MEM), fetal bovine serum (FBS), and penicillin–streptomycin solution were purchased from Gibco (Gaithersburg, MD, USA). Tartrate-resistant acid phosphatase (TRAP) staining solution and alkaline phosphatase (ALP) staining kit were obtained from Sigma-Aldrich (St. Louis, MO, USA). Alizarin red S (ARS) solution was purchased from Cyagen (Santa Clara, CA, USA). Cell Counting Kit-8 (CCK-8) was obtained from Dojindo Molecular Technology (Dojindo, Kumamoto, Japan). Recombinant mouse M-CSF and RANKL were obtained from R&D Systems (Minneapolis, MN, USA). Primary antibodies against p-ERK1/2 (9101, 1:1000), ERK1/2 (9102, 1:1000), p-JNK (4668, 1:1000), JNK (9252, 1:1000), p-p38 (4511, 1:1000), p38 (8690, 1:1000), p-p65 (3033, 1:1000), p65 (8242, 1:1000), IκBα (4814, 1:1000), NFATc1 (8032, 1:1000), c-Fos (2250, 1:1000), TRAP (15,094, 1:1000), and GAPDH (5174, 1:1000) were obtained from Cell Signaling Technology (Danvers, MA). Primary antibodies against OCN (ab93876, 1:100) were obtained from Abcam (Cambridge, MA).