Fig. 6: NHE forms pores in the cell membrane.

a Schematic of toxin-induced liposomal rupture and leakage of dye. b Colorimetric analysis of liposomes left untreated, sonicated for 5 mins at 100 amplitude (CTRL) or assessed 30 mins after treatment with individual NHE subunits (C, B, or A), heat-inactivated NHE (Heat), NHE, HBL, buffer, or bovine serum albumin (BSA). c The absorbance (OD) of residual dye following treatment as in b. d Immunoblot analysis of caspase-1, gasdermin D, and IL-1β of WT BMDMs left untreated or LPS-primed and assessed 3 h after treatment with NHE in the absence or presence of liposomes (Lipo.). e Release of IL-1β, IL-18, and TNF, and death of WT BMDMs as treated in d. f Cryo-transmission electron microscopy analysis of liposomes left untreated (n = 760) or assessed 1 h after treatment with NHE (n = 800). Quantification of membrane pores of liposomes. Arrowheads indicate pores f. Scale bar, 50 nm f. NS, not significant, **P < 0.01, ***P < 0.001, and ****P < 0.0001 (one-way ANOVA with Dunnett’s multiple-comparisons test c or student’s unpaired t test e, f). Each symbol represents an independent experiment c, e, f. Data are representative of three independent experiments (n = 3 in b–f; mean and s.e.m. in c, e, and f). Source data are provided as a Source Data file.