Human THP-1 cells were grown in RPMI-1640 media supplemented with 10% (vol/vol) FBS and 50 μM 2-mercaptoethanol (Sigma # M3148). THP-1 cells were differentiated for 2 hrs with 0.5 μM phorbol-12-myristate-13-acetate (Sigma # P8139). Differentiated THP-1 cells were primed for 18 h with ultrapure lipopolysaccharide (LPS; 1 μg/ml, Sigma #L5293). THP-1 cell culture media was changed to media without LPS and treated with ATP (5 mM, Roche, # 11162306001), or Tg (1 μM Sigma # T9033) for 4 hrs. THP-1 cells were untreated or treated with 50 μM STF-083010 for 2 hr prior to the addition of Tg (1 μM) or Tm (10 μg/ml, Sigma, # T7765) and allowed to incubate for 4 hrs. After 4 hrs the media supernatant was collected and assayed for hIL-1β by ELISA (#EH2IL1B from Thermo Scientific). Further variants of THP-1 cells used in this study were from InvivoGen: THP1-defNLRP3, deficient in NLRP3; and THP1-Null, which is a positive control line proficient for inflammasome function.
