Extended Data Fig. 1: Host-derived EVs bind LPS in vivo.

a, Plasma EV levels in wild-type (WT) mice injected with PBS or LPS (10 mg/kg) for indicated times as assessed by nanoparticle tracking analysis (NTA). b, Size distribution of EVs isolated from the plasma of WT mice injected with PBS or LPS (25 mg/kg) 1.5 h post-injection via OptiPrep density gradient method as assessed by NTA. c, Immunoblotting analysis of EVs isolated from the plasma of PBS- or LPS-injected WT mice as in (b). d, Negative staining TEM of EVs isolated from the plasma of PBS or LPS-injected WT mice as in (b). e, LPS content of the EVs isolated from WT mice injected with PBS or LPS as assessed by the LAL assay (n=4). 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 (25 mg/kg) 1.5 h post-injection as assessed by ImageStream flow cytometry (n=3). h, Immunoblotting analysis of EVs isolated by the DIC method using CD9, CD63, and CD81 antibodies-coated magnetic beads from the plasma of PBS- or FITC-LPS (25 mg/kg)-injected WT mice. i,j, LPS content of the DIC-isolated EVs from PBS- or FITC-LPS-injected mice 1.5 h post-injection as assessed by the LAL (n=3) (i) and HEK-Blue TLR4 reporter cell (n=3) (j) assays. k,l, Percentage of FITC-LPS^+ve EVs (k) and FITC histogram and mean fluorescence intensity (MFI) of EVs (l) isolated by the DIC method from mice injected with PBS or FITC-LPS as assessed by flow cytometry (n=3). Combined data from two (a) or three (e,f,g,i,j,k,l) or representative data of two (b,c,d,h) independent experiments are shown. Data are shown as mean±s.e.m (a). Each circle represents a mouse, and the horizontal lines represent mean (a, e–g and i–l). P values were determined by one-way ANOVA with Dunnett’s post-test (a) or unpaired two-tailed t-test (e–g and i– l). Scale bar, 50 nm (d).