Fig 2: Co‐staining of Anx5, lactadherin and CD41 with ApoB‐containing lipoproteins. (a and b) Scatter plots of Anx5‐Cy5, lactadherin‐FITC and CD41‐BV510 versus ApoB‐PE fluorescence depicting the degree to which markers co‐stain with ApoB in (a) a pool of PPP that was stored at −80°C after collection or (b) freshly collected PPP from one of three different individuals (representative data). Samples were either stained with Anx5 or lactadherin, as these markers both label PS. Bar plots depict the percentage of marker‐positive events that were also positive for ApoB in specifically stained samples (blue) versus isotype controls (red). Bar plots for fresh PPP depicts the mean and standard deviation (error bars) for three individuals. (c) Dot plots (large‐angle light scatter versus Anx5 or lactadherin fluorescence) of ApoB+ (red) and CD41+ (green) events demonstrate a significant overlap of these populations on these parameters. Light scatter and fluorescence intensities are denoted in arbitrary units. Anx5: Annexin V; ApoB: Apolipoprotein B; CD41: Cluster of differentiation 41/platelet membrane glycoprotein IIb‐IIIa complex; Lact: Lactadherin; LALS: Large‐angle light scatter (side scatter); PPP: Platelet‐poor plasma

Fig 3: Illustrations of the similarities and differences between the large ApoB‐containing lipoproteins (VLDL and chylomicrons) and EVs. The yellow alpha‐helical structures on the VLDL and chylomicron represent ApoB‐100 and ApoB‐48, respectively. The blue spheres on all particles refer to phospholipids while the red spheres represent PS phospholipids. The additional coloured components on the EV represent different types of membrane proteins such as tetraspannins (green). Cholesterol and other membrane components including other types of apolipoproteins on the lipoproteins have been left out for the sake of simplicity

Fig 4: Effect of detergent lysis with Triton X‐100 on ApoB‐containing lipoproteins co‐stained with Anx5, lactadherin and CD41. (a) Scatter plots of Anx5‐Cy5 and lactadherin‐FITC versus ApoB‐PE fluorescence of a representative specifically stained fresh PPP sample and its detergent lysis controls depicting the degree to which the different fluorescently labelled populations are affected by detergent lysis. Fluorescence intensities is denoted in arbitrary units. (b) Bar plots of the concentrations of different marker‐positive populations in specifically stained fresh PPP (blue) and detergent lysis controls (red). Bar plots for fresh PPP depict the mean and standard deviation (error bars) for three individuals. Anx5: Annexin V; ApoB: Apolipoprotein B; CD41: Cluster of differentiation 41/platelet membrane glycoprotein IIb‐IIIa complex; Lact: Lactadherin