Mre 3008f20

The adenosine A1 receptor agonist N⁶-cyclopentyladenosine (N6-CPA, purity > 98%) was obtained from Abcam (Shanghai, China) and dissolved in DMSO at 50 mM. The adenosine A1 receptor antagonist rolofylline (KW-3902, purity > 99%) was obtained from Tocris Bioscience (Bristol, United Kingdom) and dissolved in DMSO at 10 mM. The adenosine A2a receptor agonist CGS 21680 (purity > 98%) was obtained from Abcam and dissolved in DMSO at 10 mM. The adenosine A2a receptor antagonist SCH 58261 (purity > 99%) was obtained from Abcam and dissolved in DMSO at 10 mM. The adenosine A2b receptor agonist BAY 60-6583 (purity > 98%) was obtained from Tocris Bioscience and dissolved in DMSO at 20 mM. The adenosine A2b receptor antagonist PSB603 (purity > 98%) was obtained from Tocris Bioscience and dissolved in DMSO at 10 mM. The adenosine A3 receptor agonist piclidenoson (IB-MECA, purity > 97%) was obtained from Abcam and dissolved in DMSO at 10 mM. The adenosine A3 receptor antagonist MRE 3008F20 (purity > 98%) was obtained from Tocris Bioscience and dissolved in DMSO at 50 mM. The adenosine analog 5′-N-ethylcarboxamidoadenosine (NECA, purity > 99%) was obtained from Tocris Bioscience and dissolved in DMSO at 50 mM. All solutions were stored at –20°C and used within 1 year.

All reagents were from Thermo Fisher Scientific (Waltham, MA) unless otherwise noted. Nigericin, adenosine and MTA were from Sigma-Aldrich (St Louis, MO), while SCH442416, PSB1115 and MRE 3008F20 were from Tocris (Minneapolis, MN) and TLR ligands were from Invivogen (San Diego, CA). Anti-HMGB1 rabbit monoclonal antibody (mAb) EPR3507 was from Genetex (Irvine, CA), anti-actin mouse mAb AC-15 was from Sigma-Aldrich, anti-IL-1β mouse mAb 3ZD was from Frederick National Laboratory for Cancer Research (Frederick, MD), IL-1β ELISA antibodies were from eBioscience (San Diego, CA), TNFα, IL-6 and IL-10 ELISA antibodies were from BioLegend (San Diego, CA), FITC-conjugated anti-CD69 and APC-conjugated anti-CD86 mAb were from BD Biosciences (San Jose, CA), and anti-mouse and anti-rabbit antibodies conjugated to HRP were from Jackson Immunoresearch (West Grove, PA). Bone marrow was either a generous gift from Lisa Borghesi or obtained from C57BL/6 mice (Jackson Labs, Bar Harbor, ME) maintained in accordance with the Texas Tech Institutional Animal Care and Use Committee.