Mouse il 1β

ATDC5 murine chondrogenic cell line was obtained from RIKEN Cell Bank (Koyadai, Tsukuba, Ibaraki, Japan). Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with Ham’s F-12, Fetal Bovine Serum (FBS), antibiotics Penicillin-Streptomycin, Transferrin, Sodium Selenite, L-Glutamine, Trypsin, TRI Reagent, FNT, 3′methoxy-4′nitroflavone (3-MF), mouse IL-1β, MTT reactive, chloric acid, SDS, and Griess reactives (phosphoric acid, sulphanilamide, and alpha-naftil-etil-enamide) were purchased from Sigma Aldrich (St. Louis, MO, USA). ZK164015 was acquired from Cienytech (Santiago de Compostela, Spain). Insulin (Actrapid 100 UI/ML) was kindly donated by the Pharmacy Service of the Clinical Hospital of Santiago de Compostela. E.Z.N.A. Total RNA Kit I was bought from Omega BIO-TEK (Norcross, GA, USA). Digestion kit was purchased from Lucigen (Middelton, WI, USA). High-capacity cDNA Reverse Transcription Kit was bought from Applied Biosystems, Life Technologies (Grand Island, NY, USA). MasterMix PCR was purchased from ThermoFisher Scientific (Waltham, MA, USA). IL-6, neutrophil gelatinase-associated lipocalin (LCN2), C-C motif chemokine 2 (CCL2), COL2A1, ACAN, type X collagen (COLX), SRY-Box transcription factor 9 (SOX9), and MMP13 primers were purchased from Sigma Aldrich. Cell culture plates were purchased from ThermoFisher Scientific.

Mouse TNFα (#T539), human TNFα (#T6674), mouse IL-1β (#SRP8033), mouse IL-6 (#SRP3330) and mouse MCP-1 (#SRP3215) were purchased from Sigma-Aldrich. PD0325901 (#HY-10254), SB203580 (#HY-10256), JNK-IN-8 (#HY-13319), BI605906 (#HY-13019) and IKK16 (#HY-13687) were obtained from MedChemExpress and used in concentrations and time points indicated in figure legends.