ELISA analysis of IL-1β secretion by (a) BMDM and (b) BMDC primed with LPS and treated with the indicated stimuli (n = 3 mice per genotype). (c,d) Time course of IL-1β secretion, IL-18 secretion, and pyroptosis of peritoneal macrophages primed with LPS and treated with (c) nigericin or (d) ATP (n = 4 mice per genotype). P values: Nek7+/+ vs. Nek7+/Cu (gray); Nek7+/+ vs. Nek7Cu/Cu (magenta); Nek7+/+ vs. Nlrp3-/- (teal); Nek7Cu/Cu vs. Nlrp3-/- (blue). * P≤0.05; ** P≤0.01; *** P≤0.001; **** P≤0.0001 (unpaired, two-tailed Student’s t test). (e) TNF and (f) IL-6 secretion by peritoneal macrophages treated with the indicated stimuli (n = 4 mice per genotype). Results are representative of two independent experiments (mean and s.d. shown).