Extended Data Fig. 3: EV purification, characterization, and lipidomics details.
(a) Representative FPLC chromatograms showing A280 absorbance throughout SEC process; the collected EV fraction is shown in blue. (b) Representative NTA histograms showing size distribution detected in respective EV purifications from different sources. Lines and insets indicate mean particle diameter of 5 replicate acquisitions. (c) Phospholipid distribution in purified EVs from indicated sources analyzed by shotgun lipidomics. Shown are means of 2 biological replicates with averaged technical duplicates (saliva EVs) or 2-3 technical replicates of an individual EV preparation (other sources), each from body fluids pooled from at least 35 (semen), 9 (saliva), 6 (urine), an individual (milk) or 3 donors (blood). See Supplementary Table 3 for all lipidomics data. (d) Half-maximal inhibitory concentration (IC50) (ZIKV-MR766 MOI 0.25 on Vero E6) of liposomes containing 30 mol% PS with different individual lipid species or a biological mixture (brain PS). n = 3 independent experiments in biological triplicates, mean ± SD. (e) Nanoflow cytometry dot plots of five separate semen EV preparations stained for tetraspanins CD9/63/81 (TSPN) or PS (LA). Highlighted TSPN+ quadrants were used for quantification in Fig. 3i.
