Fig. 6

FGF10 attenuates cigarette smoke-induced endothelial apoptosis and glycocalyx repair through FGFR1 signaling. A The human pulmonary microvascular endothelial cells (hPMVECs) were treated with CSE (24 h, 2%), with or without pretreatment of FGF10 (2 h prior to CSE stimulation, 50 ng/ml) and the FGFR1 inhibitor, AZD4547 (24 h prior to CSE stimulation, 20 nM). TNF-α + SM-164 serves as positive controls (24 h, 1:500). After 24 h stimulation of 2% CSE, hPMVECs were performed for flow cytometry (n = 3). Annexin V/PI-stained cells showing apoptotic rates. FITC, fluorescein isothiocyanate; PI, propidium iodide. B Relative FGFR1 mRNA expression in hPMVECs from each group (n = 3). C Relative mRNA expression of FGFR1 and FGFR2 in hPMVECs (n = 3). D Representative western blots and corresponding group data showing expression levels of the total and phosphorylation of FGFR1. E Representative western blots and corresponding group data showing expression levels of the total and phosphorylation of ERK and AKT in hPMVECs. F Venn diagram of overlapping putative genes related both to cell apoptosis pathway and COPD, from “GO_Apoptosis_Signaling” (http://www.broadinstitute.org/gsea/index.jsp), and “New loci”, “Loci previously described” and “Candidates”. G Relative SOX9 mRNA expression in hPMVECs from each group (n = 3). H Endothelial cells were transfected with si-control and si-SOX9 for 48 h, stimulated beforehand with FGF10 (100 ng/ml) for 2 h, and then with CSE (2%) for another 24 h. Endothelial cells were performed for flow cytometry (n = 3). I Prediction of the combination site of SOX9 on the promoter region of HS6ST1. Relative HS6ST1 mRNA expression in hPMVECs from each group (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001. n.s., not significant