Iκb kinase ikk β

Recombinant murine IL-1β and recombinant murine M-CSF protein were purchased from R&D Systems (Minneapolis, MN, USA). Recombinant murine IFN-γ was purchased from Peprotech (Rocky Hill, NJ, USA). Lipopolysaccharides (LPS) were obtained from Biosharp (Beijing, China). Metformin hydrochloride was purchased from CTAO (Eugene, OSU, USA). Anti-p85-PI3K, MTOR, p-MTOR, FoxO1, p-FoxO1, AKT, p-AKT, GSK3ß, p-GSK3ß, NF-κB, p-NF-κB, IκBα, p-IκBα, IκB kinase (IKK)-β, and p-IKKα/β antibodies were supplied by Cell Signaling Technology (Beverly, MA, USA). MMP3, MMP13, INOS, and CD206 antibodies were obtained from Abcam (Cambridge, UK). Col2a1 and Sox9 antibodies were purchased from Abclonal (Wuhan, China). IL-1ß, IL-6, CD86, and F4/80 antibodies were purchased from Proteintech (Wuhan, China). GAPDH, ß-Actin, and secondary antibodies were purchased from Boster (Wuhan, China).

Recombinant murine soluble RANKL and M-CSF were purchased from R&D Systems (Minneapolis, MN, United States). JZL184 was purchased from Shanghai Lollane Biological Technology (Shanghai, China) and dissolved in DMSO (vehicle). Cell Counting Kit-8 (CCK8) was obtained from Boster Biotechnology (Wuhan, China). Antibodies against extracellular signal-regulated kinase (ERK), p-ERK, p38, p-p38, IκBα, p-IκBα, p65, p-p65, c-Jun N-terminal kinase (JNK), p-JNK, IκB kinase (IKK)-β, IKKα, p-IKKα/β, and c-Fos were purchased from Cell Signaling Technology (Beverly, MA, United States). Antibodies targeting tartrate-resistant acid phosphatase (TRAP), MMP-9, CTSK, and NFATc1 were acquired from Abcam (Cambridge, United Kingdom). The TRAP staining kit was purchased from Sigma-Aldrich (St. Louis, MO, United States). The Osteo-Assay Surface assay was obtained from Corning Life Sciences Inc., (Corning, NY, United States). Basal culture medium was obtained from Invitrogen (Carlsbad, CA, United States).

Recombinant mouse IL-1β was purchased from R&D Systems (Minneapolis, MN, United States). Lipofectamine 3000 reagent was obtained from Invitrogen (Waltham, MA, United States). Anti-SHP2, β-catenin, p38, p-p38, extracellular signal-regulated kinase (ERK), p-ERK, c-Jun N-terminal kinase (JNK), p-JNK, p65, p-p65, IκBα, p-IκBα, IκB kinase (IKK)-β and p-IKKα/β antibodies were supplied by Cell Signaling Technology (Beverly, MA, United States). MMP3, MMP13 and p-β-catenin(Y142) antibodies were obtained from Abcam (Cambridge, United Kingdom). p-β-catenin(S33/37/141), COL2A1, and AGGRECAN antibodies were purchased from Abclonal (Wuhan, China). GAPDH antibody was purchased from Boster (Wuhan, China) and secondary antibodies were obtained from the Jackson lab.