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. Author manuscript; available in PMC: 2024 Jun 1.
Published in final edited form as: Nat Cell Biol. 2023 Nov 16;25(12):1860–1872. doi: 10.1038/s41556-023-01269-8

Fig. 4: EV-mediated cytosolic LPS delivery activates the noncanonical inflammasome.

Fig. 4:

a,b, GSDMD (FL, full length; NT, N-terminal domain), caspase-11 and β-actin in the lysates (a) and cell death (LDH release; b) of IFN-γ-primed (10 ng/ml) WT BMDMs stimulated with medium, varying doses of PBS-EVs or LPS-EVs (6.25–50 μg/ml), LPS or LPS transfection for 16 h. c,d, GSDMD, caspase-11, and GAPDH in the lysates (c) and cell death (LDH release; d) of IFN-γ-primed (10 ng/ml) WT and Casp11−/− BMDMs stimulated as indicated for 16 h. e, GSDMD, caspase-11, and β-actin in the lysates of spleen and liver of poly(I:C)-primed (150 μg) WT mice injected i.p with SEC-isolated PBS-EVs or LPS-EVs for 15 h (n=3). f–i, IL-18 (f), IL-1β (g) , IL-1α (h) and galectin-1 (i) in the plasma of poly(I:C)-primed WT mice injected with SEC-isolated PBS-EVs or LPS-EVs for 15 h (n=9). j,k, IL-18 and IL-1β in the plasma of poly(I:C)-primed WT mice injected with UC-isolated PBS-EVs or LPS-EVs for 6 h (n=7). l,m, IL-18 and IL-1β in the plasma of poly(I:C)-primed WT mice administered with UC-isolated EVs from LPS-injected WT and Casp1/11−/− mice (n=4). n, GSDMD, caspase-11, and β-actin in the lysates of liver and spleen of poly(I:C)-primed WT and Casp11−/− mice injected with SEC-isolated LPS-EVs for 15 h (n=3). o,p, IL-18 and IL-1β in the plasma of WT and Casp11−/− mice treated as described in (n) (n=6). q, Survival of poly(I:C)-primed WT and Casp11−/− mice injected with SEC-isolated LPS-EVs (n=9). Combined data from two (j–m,o,p) or three (b,d,f,g,h,i,q) independent experiments or representative data of two (a,c,n) or three (e) experiments are shown. Each circle represents a mouse and the horizontal lines represent the mean (f-m,o,p). Each lane represents a mouse (e,n). Data are presented as mean±s.e.m (b,d). P values were determined by unpaired two-tailed t-test (f-m,o,p), one-way ANOVA with Dunnett’s post-test (b), two-way ANOVA with Sidak’s post-test (d) or Mantel-Cox test (q).