Figure 1.

NLRP3 expression, priming, and/or activation triggers pyroptosis in reconstituted U937 depending on the NLRP3 variants. (A–G) NLRP3-deficient U937 cells reconstituted with doxycycline-inducible NLRP3 variants were treated with doxycycline (1 μg/ml, 3 h), LPS (40 ng/ml, 2 h), and nigericin (15 μg/ml) before cell death was monitored by PI incorporation over time quantified by time-lapse high content microscopy. (B) Example of microscopy images for WT and K568N (60 min, objective 10×, scale bar 100 μm). (C) K568N (group#5). (D) E311K (group#4). (E) A439V (group#3). (F) R168Q (group#2). (G) T952M (group#1). Means of duplicates and 1 SD (left panel) and means of AUC of duplicates and 1 SD (right panel) for cells expressing NLRP3 variants (full square) and WT (open circle) are represented. One experiment done in duplicates representative of two to eight independent experiments is shown (as indicated for each variant on the right side). Results obtained with one variant typical of each functional group are represented. Results obtained with all tested variants are presented in Fig. S2, B–F. Statistical analysis including all independent experiments are represented in Fig. 2.