Extracellular vesicle cross-talk between pulmonary artery smooth muscle cells and endothelium during excessive TGF-β signalling: implications for PAH vascular remodelling

摘要

Characterisation of EVs released by HPASMC in vitro. , Schematic representation of the approach used to analyse EVs-mediated HPASMC-to-HPAECs communication. , 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. , Immunocytochemistry showing CD63+ EVs from HPASMCs in cell culture (representative image from  = 4 independent experiments). , 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. , 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