Fig. 3

Stimulation of TGF-β signalling triggers fibrotic phenotype and downregulates inflammatory pathways on HPASMCs. a, Control, TGF-β1-treated and BMP4-treated HPASMCs were analysed by RNA-Seq. PCA analysis of HPASMCs showed a greater effect of TGF-β1 than BMP4, as these HPASMCs were separated from the rest by the first principal component, accounting for 69% of the variance. b, Heatmap showing the activation of specific downstream markers of TGF-β1 and BMP4 pathways. c, GO analysis for biological process of the differential transcripts in TGF-β1-treated HPASMCs provided significant terms relevant to PAH: cell migration (yellow), proliferation (brown) and inflammation (grey), as well as with cell communication (pink). d, Interactome analysis of the top differential RNAs in TGF-β1-treated HPASMCs. Most relevant clusters are highlighted: Inflammation, ECM remodelling/cell adhesion, serotonin signalling and signal transduction. e, Heatmap showing the inflammatory markers downregulated in response to activation of TGF-β signalling in HPASMCs. Treatment of HPASMC with TGF-β1 or BMP4 (both 10 ng/ml) was performed for 48 h and renewed every 24 h. Protein interaction analyses were performed with STRING and GO enrichment using DAVID software. PCA = principal component analysis, GO = Gene Ontology