Figure 1. Alum and LLOMe trigger Nlrp3-independent cell death and are poor inducers of IL-1β release. (A and B) BALB/c macrophages were primed with 250 ng/ml LPS in the presence or absence of 100 μM CA-074-Me for 2 h. Macrophages were then exposed to increasing concentrations of LLOMe, ATP or nigericin for 4 h or alum for 6 h, in the presence or absence of 40 μM Boc-D-CMK. IL-1β release was assessed by ELISA (A), and cell death by CytotoxOne LDH activity assays (B) from supernatants. (C) Wild type and Nalp3-, Ipaf- and Asc-deficient C57BL/6 macrophages were primed for 2 h with 250 ng/ml of LPS in the presence or absence of 100 μM CA-074-Me. Cells were then treated with 150 μg/ml alum, 2.5 mM LLOMe or with varying doses of ATP and nigericin for 3 h. Plasma membrane impairment was determined by propidium iodide exclusion assays. All measurements were taken in triplicate from representative experiments.