Fig. 1.

Characterisation of EVs released by HPASMC in vitro. a, Schematic representation of the approach used to analyse EVs-mediated HPASMC-to-HPAECs communication. b, Quantification of the downstream gene Serpine1 by qRT-PCR shows specific activation with TGF-β1. Id1 was highly activated by BMP4 and mildly by TGF-β1 due to these signalling pathways´ crosstalk, as expected. c, Immunocytochemistry showing CD63+ EVs from HPASMCs in cell culture (representative image from n = 4 independent experiments). d, Identity of HPASMC-EVs isolated by ultracentrifugation was confirmed by expression of CD63 marker, as opposed to their donor HPASMCs. Histone 3 used as a control for cell contamination was only detectable in HPASMCs. GAPDH was used as HPASMC loading control. e, The HPASMC-EVs isolated were analysed by Nano Tracking Analysis (NTA) which showed EVs ranging 80–500 nm with a highest peak at 150 nm