Fig. 6: Analysis of HMVECs secretome induced by TNF-α.

HMVECs were treated for 24 h and 48 h with 100 ng/ml of TNF-α and supernatants were analysed by mass spectrometry. A The Venn diagram presents proteins detected in HMVECs secretome treated or not with TNF-α. In total, 602 proteins were detected whose 15 only in HMVECs secretome, 80 only in HMVEC’s secretome treated with TNF-α and 507 were common to both. B The Venn diagram depicting total protein distribution, 20 proteins were not significantly different between subpopulations, 36 and 409 were significantly elevated in the endothelial and mesenchymal subpopulations, respectively, using a 1.5 cut-off for significant differences. Heatmap of proteins differentially expressed between Ctrl and TNF-α at 24 h and 48 h and between TNF-α 24 h and TNF-α 48 h. Results were obtained with three technical replicates. C, D TNF-α induces an increase of secretory capacity of HMVECs as revealed by the sum of abundance intensity (iBAQ) for all identified proteins (C) and an increase of COL1A1 secretion (D). E, F All proteins detected at 48 h of TNF-α treatment were analysed with IPA. Pathways (E) and functions (F) significantly affected are represented by the graphs. G mRNA expression levels of TNSF12 and TNFRSF12 in PDXs. HMVECs human microvascular endothelial cells, TNF-α tumour necrosis factor-α, Ctrl control, COL1A1 collagen type I α1, TNFSF12 tumour necrosis factor ligand superfamily member 12, TNFRSF12 tumour necrosis factor receptor superfamily member 12, PDXs patient-derived xenografts.