Recombinant murine m csf

Purified Schisandrin B (purity: 99.99%), extracted from Schisandra chinensis, was obtained from MedChemExpress (Monmouth Junction, NJ, USA). Recombinant murine M-CSF and RANKL were purchased from R&D Systems (Minneapolis, MN, USA). Antibodies against NFATc1, c-FOS, P38, JNK, ERK, P65, IκBα, p-P38, p-JNK, p-ERK, p-P65, and p-IκBα were obtained from Cell Signaling Technology (Beverly, MA, USA). Antibody targeting TRAP was purchased from Abcam (Cambridge, MA, USA). Antibodies against MMP9, Nrf2, HO1, NQO1, and β-Actin were from Proteintech Group (Wuhan, Hubei, China). Trap staining kit and other reagents were purchased from Sigma-Aldrich (St. Louis, MO, USA).

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).

Magnoflorine [C₂₀H₂₄NO₄; MW, 342.41; purity, ≥98% (Meilun, Dalian, China)] was dissolved in dimethyl sulfoxide. Recombinant murine M-CSF and RANKL were from R&D Systems (Minneapolis, MN, USA). Fetal bovine serum and alpha-modified minimal essential medium (α-MEM) were supplied by Gibco-BRL (Sydney, Australia). The staining kit for tartrate-resistant acid phosphatase (TRAP) was obtained from Sigma-Aldrich (St. Louis, MO). The assay for cell viability, cell counting kit-8 (CCK-8), was from Dojindo Molecular Technology (Japan). Primary antibodies targeting extracellular signal-regulated kinase (ERK) and its phosphorylated form (phospho-ERK; Thr202/Tyr204), p38 and phospho-p38 (Thr180/Tyr182), TAK1 and phospho-TAK1, c-Jun N-terminal kinase (JNK) and phospho-JNK (Thr183/Tyr185), NF-κB (p65) and phospho-NF-κB, NF-κB inhibitor alpha (IκBα) and phospho-IκBα, c-Fos, β-actin and histone H3 (as internal controls), and appropriate secondaries conjugated to fluorescent dyes were from Cell Signaling Technology (Cambridge, MA). The primary antibody recognizing nuclear factor of activated T cells, cytoplasmic 1 (NFATc1) was supplied by Absin Bioscience Inc. (Shanghai, China). The Prime Script RT reagent kit and SYBR^® Premix Ex Taq™ II were from Takara Biotechnology (Otsu, Japan).

Fetal bovine serum (FBS) and α-minimum essential medium (α-MEM) were obtained from Gibco BRL (Grand Island, NY, USA). Recombinant murine M-CSF and RANKL were purchased from R&D Systems (Minneapolis, MN, USA). The acid phosphatase, leukocyte (tartrate-resistant acid phosphatase: TRAP) kit, methylthiazolyldiphenyl-tetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), and all other reagents were purchased from Sigma–Aldrich (St. Louis, MO, USA). (-)-Tubaic acid (4-hydroxy-2-(1-methylethenyl)-2,3-dihydrobenzofuran-5-carboxylic acid) was isolated by the thermal transformation of rotenone, and the isolation process is described later in this section.