Z levd fmk

Human IL-1β ELISA Set II, human TNF ELISA Set, human IL-6 ELISA Set and mouse IL-1β ELISA Set (BD OptEIA). THP1 cells (1 × 10⁶ cells per well in a six-well plate) were transduced with scramble or shSERPINB1 lentivirus for 48 h and primed with 1 μg ml⁻¹ LPS (0127:B8, Sigma) for 12 h. For NLRP3 inflammasome activation, THP1 cells were differentiated with 100 ng ml⁻¹ phorbol 12-myristate 13-acetate (Calbiochem) for 72 h, transduced by scramble or shSERPINB1 lentivirus for 48 h, and primed with LPS (0.5–1 μg ml⁻¹) overnight. Cells were washed by PBS and stimulated with control media for 3 h, 2 μM nigericin (Sigma) for 3 h, 10 μg ml⁻¹ muramyl dipeptide (Sigma) for 6 h, 5 mM ATP (Sigma) for 1 h, 2.5 μg ml⁻¹ flagellin (Invivogen) for 6 h, or 1 μg ml⁻¹ poly(dA:dT)/LyoVec (Invivogen) for 6 h. For caspase inhibitor treatment, THP1 cells were transduced with scramble or shSERPINB1 lentivirus for 48 h, treated with indicated caspase inhibitors (20 μM) 1 h before LPS (1 μg ml⁻¹, 12 h) priming. Caspase inhibitors include z-YVAD-FMK (Santa Cruz), z-LEVD-FMK (BioVision), z-WEHD-FMK (BioVision), z-DEVD-FMK (Santa Cruz) and z-VAD-FMK (Santa Cruz).