Disuccinimidyl suberate dss 21655

Phorbol 12-myristate 13-acetate (PMA, P8139), lipopolysaccharides from E. coli O111:B4 (L2630), adenosine triphosphate (ATP, A2383), potassium chloride (KCl, P5405) MG132 (C2211), folic acid (F876), allopurinol (A8003), and dexamethasone (D2915) were purchased from Sigma‒Aldrich (St. Louis, MO, USA). Recombinant human TNFα (210-TA), human M-CSF (216-MC), mouse IL-1β (401-ML), and mouse TNFα (410-MT) were purchased from R&D Systems (Minneapolis, MN, USA). Monosodium urate crystals (MSU crystals, tlrl-msu), Salmonella flagellin (FLS-ST, tlrl-stfla), and poly (dA:dT) (tlrl-patn) were obtained from InvivoGen (San Diego, CA, USA). Complete Freund’s adjuvant (CFA, 7001) and incomplete Freund’s adjuvant (IFA, 7002) were obtained from Chondrex (Redmond, WA, USA). Colchicine (AG-CN2-0048) was purchased from AdipoGen (San Diego, CA, USA). Recombinant mouse M-CSF (315-02) was purchased from PeproTech (Rocky Hill, NJ, USA). Recombinant human IL-1β (RIL1B1) and disuccinimidyl suberate (DSS, 21655) were purchased from Thermo Fisher Scientific (Cleveland, OH, USA). Human NCOA6 siRNA (sc-61401), control siRNA (sc-37007), and febuxostat (144060-53-7) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). ON-TARGETplus mouse Ncoa6 siRNA (L-041129-00-0005) and ON-TARGETplus nontargeting control siRNA (D-001810-10-05) were purchased from Dharmacon (Lafayette, CO, USA).

Mice with LoxP sites flanking both sides of exon 4 of Bmal1 (Bmal1LoxP/LoxP, gifted from the lab of Dr. Christopher A. Bradfield) were crossed with mice containing a lysozyme M activated CRE recombinase (Lyz2Cre, Jackson Labs, #004781) to generate mice with the Bmal1 gene excised specifically in the myeloid lineage (Bmal1LoxP/LoxPLyz2Cre) i.e. Bmal1^-/-. Mice with a lysozyme M activated CRE recombinase (Lyz2Cre) were used as controls i.e. Bmal1^+/+. Mice used for experiments were both male and female, aged between 8-12 weeks. These mice were bred and maintained in specific pathogen free conditions in the Comparative Medicine Unit (CMU), Trinity College Dublin. All mice were maintained in line with European Union and Irish Health Products Regulatory Authority (HPRA) regulations. Experiments were carried out under HPRA license and with ethical approval from Trinity College Dublin (TCD) bioethics committee and Royal College of Surgeons in Ireland (RCSI) research ethics committee. Lipopolysaccharides from Escherichia coli O55:B5 (L2880), dimethyl malonate (136441), and 2-deoxyglucose (H0887) were purchased from Sigma. Disuccinimidyl suberate (DSS) (21655) was purchased from Thermo Scientific. Stattic (AB120952) was purchased from Abcam. DASA-58 (HY-19330) was purchased from MedChemExpress.