Extended Data Fig. 2: NINJ1 is essential for pyroptosis-related PMR.
From: NINJ1 mediates plasma membrane rupture during lytic cell death
a, Immunoblot of NINJ1 and GSDMD in BMDM lysates. b, Left, release of DD-150 in live-cell imaging analysis of iMACs after LPS electroporation over a 16-h time course. Right, immunoblot of NINJ1 in Ninj1–/– iMACs reconstituted with NINJ1. Data are means (circles) ± s.d. (shaded area) of three individual replicates. c, Immunoblot of GSDMD, GSDMD-NT and NINJ1 in supernatant and extract from nigericin-stimulated primed BMDMs. Actin is from a separate blot. d, LDH or IL-18 release from BMDMs stimulated as in Fig. 2d. e, IL-6 or TNF production from BMDMs stimulated with Pam3CSK4 (TLR2) or extracellular LPS (TLR4). f, Lipid composition of primed BMDMs. Lipids profiled are diacylglycerol (DAG), dihydroceramide (DCER), hexosylceramide (HCER), lactosylceramide (LCER), lysophosphatidylcholine (LPC), lysophosphatidylethanolamine (LPE), phosphatidylcholine (PC), phosphatidylethanolamine (PE), sphingomyelin (SM), triacylglycerol (TAG), cholesteryl ester (CE), and ceramide (CER). g, RNA-seq of primed BMDMs. h, Time-lapse imaging analysis of mitochondrial membrane potential as measured by tetramethylrhodamine methyl ester perchlorate (TMRM) in primed BMDMs following LPS electroporation. Data are means (circles) ± s.d. (shaded area) of three individual replicates. i, Single-cell time-lapse imaging analysis of TMRM and DD-150 release in primed wild-type BMDMs after LPS electroporation. Time point ‘0’ corresponds to the point of maximal TMRM decline for each cell. j, Silver staining of proteins in culture supernatant of nigericin-stimulated primed BMDMs. k, Kaplan–Meier survival plots for mice challenged with 54 mg kg−1 LPS. P values were calculated by a two-sided Gehan–Breslow–Wilcoxon test. Unless otherwise specified, data are means (bars) of at least three individual replicates (circles). n = 2 per genotype. For gel source data, see Supplementary Fig. 1.
