BAL-0028 inhibits human NLRP3 in mouse cells . Related to Fig. 3. (A and B) Comparison of BAL-0028 and MCC950 in LDH release assays from (A) WT 129S6 iBMDM and (B) 129S6-human promoter NLRP3 iBMDM cells stimulated with LPS and nigericin. Graph symbols show average LDH values ± SEM from N = 3 independent experiments (indicated by different symbols) performed in triplicate. (C and D) Comparison of BAL-0028 and MCC950 in IL-1β release assays from (C) WT 129S6 and (D) 129S6-human promoter NLRP3 primary peritoneal macrophages stimulated with LPS and nigericin. Graph symbols show average IL-1β values relative to vehicle control ± SD from one experiment performed in duplicate. IC50 curves were fitted by nonlinear regression analysis. (E) Multiple sequence alignment of human, African green monkey (AGM), cynomolgus monkey (CYNO), dog, rabbit, mouse, and rat NLRP3 protein sequences restricted to the corresponding sequence of amino acids 131–694 in the human NACHT domain construct shown below the alignment.