Fig. 2

HPASMC-EVs show a specific transcriptome and are enriched in Zeb1, GDF11 and TGF-β3. Data from the low-input RNA-Seq was analysed to examine the cargoes sorted into HPASMC-EVs. Enrichment analysis was performed consisting in the quantification of cargoes with higher expression in EVs compared to their donor HPASMCs (pvalue < 0.05 and FC > 2). a, Proportion of coding/non-coding RNAs detected in HPASMC and their EVs. b, Volcano plot showing the transcripts depleted or enriched in EVs compared to donor HPASMCs (pvalue < 0.05 and FC < − 2 or FC > 2). Among the enriched we found: Zeb1, GDF11 and TGF-β3. c, Validation by qRT-PCR of the enrichment of GDF11, TGF-β3 and Zeb1 in HPASMC-EVs (n = 3 biological replicates, data are presented as mean ± SEM. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001 (Student’s t-test). d, Immunocytochemistry of Zeb1 and CD63 proteins showing co-localisation within the cytoplasm of HPASMCs. e, Western Blot showing a specific expression of Zeb1 protein in EVs. Scale bars = 10 μm. Protein interaction analyses were performed with STRING and GO enrichment using DAVID software. PCA = principal component analysis, GO = Gene Ontology