Fig. 4

Flow cytometry of plasma extracellular vesicles. Representative gating strategy of plasma derived lEVs showing first counting beads as reference (left graph). Selection of events positive for tetraspanin markers (CD9, CD63 and CD81) was followed by selection of events based on a size gate established with silica beads, additionally showing sensitivity to Triton X and an unstained control (A). Bar charts showing absolute number of lEVs per microliter comparing pre- and postprandial timepoints based on measurement with counting beads (B). t-distributed stochastic neighbor embedding (t-SNE) plots showing the mean fluorescence intensity of CD31, CD106, CD44, and CD324 in a total of 80,400 positive events (based on CD9+ , CD63+ CD81+ events and size) in each pre- and postprandial states (C). Violin plots showing the frequency of positive events for CD31 and CD106 (endothelial markers) as well as CD44 and CD324 (epithelial markers) in pre- vs postprandial states (D). n = 12, all samples were measured in technical duplicates. Statistical analyses were performed using paired t-test. P values: * for p ≤ 0.05; ** for p ≤ 0.001; *** for p ≤ 0.0001.).