ELISAs were conducted as previously described [25 (link)]. Supernatants from healthy and rheumatoid arthritis donor cell cultures treated with PLX51107 (50 nM, 100 nM, 200 nM, 250 nM) or the vehicle, dimethyl sulfoxide (DMSO, 250 nM), were collected after 48 h using the Human TNFα DuoSET ELISA kit (R&D Systems, Minneapolis, MN, USA) utilizing manufacturer provided instructions. To measure FcγR-mediated TNFα production, both healthy- and RA-donor cell cultures were treated with 350 μg/mL heat aggregated IgG (ΔIgG) in the presence of 1 μL of polymyxin B for 24 h.
Free full text: Click here
