Phosphor tak1

Antibodies against GAPDH, COL2A1, ACAN, ADAMTS4, ADAMTS5, MMP9, MMP13, TNF-α, NOX1, NOX2, iNOS, COX2, and NLRP3 were purchased from Abcam Inc. Catalase, SOD1, BAX, BCL2, Caspase3, Cleaved-Caspase3, TAK1/NF-κB signaling proteins (TAK1, phosphor-TAK1, P65, phosphor-P65, IκBα, and phosphor-IκBα), and IgG secondary antibodies were purchased from Cell Signaling Technology Inc. Antibody against GAPDH was from Proteintech Group Inc. Aloin reagent was from MedChemExpress, and human TNF-α was from R&D Systems. Abbreviations and descriptions were shown in Supplement Table 1.

Primary antibodies for iNOS, COX-2, β-actin, PARP, phospho-PI3K p85, PI3K, Akt, phospho-Akt, NF-κB, phospho-NF-κB, IκB-α, phospho-IκB-α, IKKβ, phospho-IKKα/β, phospho-p38, p38^MAPK, phospho-JNK, JNK, phospho-ERK1/2, ERK1/2, MKK4, MKK6, MyD88, IRAK1, TRAF6, phosphor-TAK1, TAK1, phospho-c-fos, phosphor-c-jun, phosphor-ATF2, IL-1β, TNFα, and horseradish peroxidase-conjugated secondary antibody were from Cell Signaling Technology (Danvers, MA, USA). SB203580, SP600125, and PD98059 were from Sigma-Aldrich (St. Louis, MO). Consensus oligonucleotides for NF-κB and AP-1 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). γ⁻³² P-Labeled ATP was purchased from ICN (Costa Mesa, CA). A prostaglandin E₂ EIA kit was from Enzo life Sciences (Ann Arbor, MI, USA). Easy Blue™ RNA extraction kit was from iNtRON, Korea. Millipore MILLIPLEX™ Mouse Cytokine/Chemokine enzyme-linked immunosorbent assay kit was from Millipore Corp. (St. Charles, MO). All materials, equipment, and biotinylated marker proteins for gel electrophoresis were from Bio-Rad. All other chemicals were purchased from Sigma-Aldrich (St. Louis, MO) unless otherwise stated. Torilin (98%) was prepared as previously indicated in [26 (link)] and dissolved in dimethyl sulfoxide and freshly diluted in culture media for all experiments.