Extended Data Fig. 2: Cell-type specific sEV enrichment of miRNAs and sEV versus cell housekeeping miRNAs. Effect of percentage of CG and Gibbs free energy (ΔG) in miRNA sorting or cellular retention.

a) Normalized gene expression of representative miRNAs showing cell-type specific sEV enrichment: miR-696 (BAT), miR-770-5p (C2C12), miR-1927 (3T3-L1), miR-1931 (AML12) and miR-718 (SVEC). Average Ct from the whole miRNA profile was used for normalization for each sample (n=3-4). *P≤0.05 (Limma t-test). b) Normalized gene expression of two representative miRNAs (miR-138-5p and miR-501-5p) showing similar sEV and cellular expression for each cell type. Average Ct from the whole miRNA profile was used for normalization for each sample (n=3-4). c) Percentage of CG content in the sequence of the miRNAs: those sorted into the sEV in all cells are shown in red (n=13); those sorted into sEV in 3 or 4 of the five cell types are shown in green (n=90); those not enriched in either sEV or cells are shown in black (n=109); those retained in 3 or 4 cell types are shown in pink (n=97); and those retained in the cell bodies of all cell types are shown in blue (n=43). d) The calculated Gibbs free energy (ΔG) for each of the miRNA in each of the five groups described in panel c. For c, d lines indicate mean value, **P≤0.01, *** P≤0.001 between the indicated group and the other four groups (ANOVA followed by Bonferroni post-hoc test in c, d). Data are expressed as mean ± SEM.