Fig. 6

EV-mediated transfer of mRNA from HPASMC to HPAECs is enhanced by TGF-β1 and IL-1β. a, Our primary cell Cre-loxP system was used to analyse the alteration of uptake and translation of EV-mRNA by activated HPAECs in vitro. We analysed 5 cytokines related to PAH: TGF-β1 (5 ng/ml), BMP4 (5 ng/ml), IL-1β (5 ng/ml), TNF-α (1 ng/ml) and VEGFA (20 ng/ml). Stimuli were renewed every 48 h during the 7 days protocol. b, Quantification of Cre-induced recombination by FACS showed significant differences in TGF-β1 and IL-1β treated HPASMC Cre+: HPAECs Rep+ co-cultures as compared to the untreated. c, NTA showed no alteration of release of EVs/cell to supernatant in HPASMCs treated with TGF-β1 and IL-1β (both 10 ng/ml, 48 h and renewed every 24 h). d, Schematic representation of the biological relevance of the achieved results. Data are presented as mean ± SEM. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001 (Student’s t-test). All procedures were carried out with n = 3 biological replicates. Scale bars 4x = 300 μm; 10x = 120 μm