Inflammasome Activation in Human Cells

Human IL-1α, IL-1β, and IFNγ were purchased from Peprotech and used at 10 ng/ml unless indicated otherwise. IL-1α and IL-1β were used interchangeably with the same results. Human IL-1ra was purchased from Peprotech and was used at 1 µg/ml. Poly IC was purchased from Sigma and used at 10 µg/ml. LPS from Escherichia coli strain 0111:B4 was purchased from Sigma and was used at 100 ng/ml. Cells were treated for 6 h for Q-PCR and 24 h for ELISA, unless indicated otherwise. Cell treatment with inflammasome activators was performed following the published protocols [41] (link)–[43] (link). ATP (adenosine 5′-triphosphate disodium salt) was purchased from Sigma and was used at 5 mM. ATP was added to cultures 30 min before cell harvest. Nigericin sodium salt was purchased from Sigma and was used at 20 µM. Nigericin was added to culture 1 h before cell harvest. Lactacystin was purchased from Santa Cruz Biotechnology and was added to culture 10 min prior to cell stimulation.