Extended Data Fig. 6: Isolation of sEV and NV using an additional step of size exclusion chromatography.

a) Diagram of the isolation method used to obtain cellular, sEV-p100 pellet, sEV-SEC and NV-SEC samples using two rounds of ultracentrifugation followed by size exclusion chromatography (SEC). (n=4). b) NTA analysis for EV concentration (top graph) of the 30 fractions obtained from SEC. These fractions were pooled in pairs in some cases and concentrated using Amicon centrifugal 3 KDa filter prior assessing protein concentration (bottom graph). c) Immunoblot for classical exosomal markers CD63 and CD9 for the concentrated fractions shown in b, bottom graph. d) PCA plot for the miRNA profile from the cells, sEV-p100, sEV-SEC and NV-SEC samples. e) Heatmap of the top differentially expressed miRNAs among sEV-p100, sEV-SEC and NV-SEC. High expression is shown in red and low expression in blue. f) Pearson correlation between averaged normalized miRNA expression levels in sEV-p100 and sEV-SEC. Expression levels were first normalized to average ct of each sample. g) Motifs found overrepresented in sEV-SEC enriched-miRNAs compared to cellular-enriched miRNAs.