Fig. 1

ERAP1 deficient macrophages have increased IL-1β, IL-18, and capsase-1 activation upon inflammasomes activation. Bone marrow-derived macrophages (5 × 10⁵ cells) were plated into 24-well plates. Cells were primed with LPS (20 ng/mL) for 16 h, and then stimulated for another 24 h with alum (1 mg/mL), nigericin (10 ug/mL), MDP (10 ug/mL), flagellin (250 ng/mL), and poly(dA:dT) (8 μM). Production of IL-1β (A–C) and IL-18 (D–F) in cell supernatant was performed. Caspase-1 activity (G) was evaluated using FLICA flow-based assay. Intracellular pro-IL1β was detected by flow cytometry using Cytek Aurora with various conditions. Baseline mock levels (H) as well as percent reduction of pro-IL-1β with priming and nigericin stimulation was compared in WT and ERAP1^−/− (I). Cells were plated in quadruplicate. Data are expressed as means ± SEM. These figures are representative of four independent experiments