XIAP-deficient BMDMs hypersecrete IL-1β in response to TLR agonists without the requirement for a second activation signal and in response to TNF-α. (A) Stimulation via various TLR agonists or TNF-α alone leads to IL-1β secretion in XIAP-deficient BMDMs but not WT BMDMs unless WT BMDMs are also stimulated concurrently with nigericin. (B) Comparable levels of TNF-α production indicate that BMDMs from XIAP-deficient or WT mice were stimulated similarly. (C) Pro–IL-1β and NLRP3 were upregulated in both WT and XIAP KO cells upon priming. Horizontal black lines on Western blot images indicate that bands of different molecular weights from the original probe and subsequent reprobes with antibodies against the indicated proteins were cropped together. Vertical black lines indicate cropping to remove an intervening lane between the “No treatment” and either “CpG” or “TNF- α” lanes (the cropped lane contained lysate from quercetin-treated cells). (D) Comparable cell loss in both XIAP and WT BMDMs except following TNF-α. Symbols used: n.s for not significant, * for p≤0.05, ** for p≤0.01, and *** for p≤0.001.