Skip to main content
. 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. 2: Host-derived EVs bind LPS in vivo.

Fig. 2:

a, Nanoparticle tracking analysis (NTA) of the size distribution of EVs isolated from the plasma of wild-type (WT) mice injected with PBS or LPS (25 mg/kg) 1.5 h post-injection via the IZON qEV column-based size exclusion chromatography (SEC). b, Immunoblotting analysis of EVs isolated as described above for CD9 and CD63. c, Negative staining TEM of EVs isolated from the plasma of PBS- or LPS-injected WT mice as described above. d,e, LPS content of the EVs isolated from PBS- or FITC-LPS-injected mice by the SEC method 1.5 h post-injection as assessed by the LAL (d; n=10) and HEK-Blue TLR4 reporter cell (e; n=7) assays. f,g, Percentage of FITC-LPS+ve EVs (f) and FITC histogram and mean fluorescence intensity (MFI) of EVs (g) isolated from mice injected with PBS or FITC-LPS as assessed by ImageStream analysis (n=6). h, TEM of EVs isolated as described above and stained with gold-conjugated anti-FITC antibody. Arrows indicate LPS. i, LPS content of the EVs isolated from WT and Casp11−/− mice injected with PBS or FITC-LPS (25 mg/kg) 1.5 h post-injection via SEC as assessed by the LAL assay (n=4). j,k, Percentage of FITC-LPS+ve EVs (j) and FITC MFI of EVs (k) isolated from WT and Casp11−/− mice injected with PBS or FITC-LPS 1.5 h post-injection via SEC as assessed by ImageStream flow cytometry (n=3). Combined data from three (j,k) four (i), six (f,g) or seven (d,e) independent experiments or data from one experiment representative of two (a–c,h) are shown. Each circle represents a mouse, and the horizontal lines represent the mean (d–g,i–k). P values were determined by unpaired two-tailed t-test (d–g) or one-way ANOVA with Dunnett’s post-test (i,j,k). Scale bar, 50 nm (c,h).