Fig 1: VEGF expression in the ARPE-19 cells and ARPE-19-isolated exosomes. (a) The VEGFA protein levels were measured by an ELISA assay in the ARPE-19 cells. (b, c) The VEGF protein levels were measured by a western blot in the exosomes. (d) Flotillin-1 levels. (e) ANXA2 levels. The experiments were repeated three times to ensure the consistency of the results. The values are expressed as (pg/ml) the mean ± S.E.M. (N = 3 repeats). The significance levels are expressed as P < 0.05 (*) versus CNT, P < 0.01 (**) versus CNT P < 0.01 (°) versus HG. CNT = standard medium; HG = 35 mM D-glucose; D.U. = Densitometric Units.
Fig 2: Phenotypic adaptation of endothelial cells in angiogenic tumoursA. Sorted ECs (eGFP+CD31+) of normal brain (NB), invasive (P8), intermediate (P3) and angiogenic (P13) glioblastoma xenografts were used for gene expression analysis. Cluster analysis of gene expression profiles indicates that xenograft-derived ECs are more closely related to each other and differ from normal brain ECs. B. Revigo summary of main GO terms (DAVID® database) up-regulated in ECs within angiogenic tumours compared to normal brain (FDR < 0.01, any fold change). Color scale represents the log10 p-value. See Suppl. Table 7 for complete list of GO terms. C. Venn diagram showing the comparison of up-regulated DEGs (see Suppl. Table 8 for detailed list of DEGs). D.-E.. Immunohistochemistry confirming increased expression of Angiopoietin 2 (Angpt2), D and Thrombospondin 1 (Thbs1), E in ECs of the intermediate and angiogenic tumours (D: scale bar 30µm; E: scale bar 20µm). Note that increased expression is also visible in the tumour cells of the angiogenic phenotype. F. Integrative protein-protein interaction analysis using genes up-regulated in tumour cells and ECs of the angiogenic phenotype (selected tumour list: cell membrane and extracellular matrix proteins, P13vsP8 cells, FDR < 0.01, FC > = 2; selected EC list: cell membrane and extracellular matrix proteins P13vsNB, FDR < 0.01, FC > 1;). The network shows direct protein-to-protein interactions between tumour-specific modules (‘grey’) and EC-specific modules (‘green’), as well as indirect interactions via putative protein partners (‘yellow’). Genes expressed by both modules are indicated in the ‘red’ module. Number of genes per module is displayed in brackets. G. Selected network of protein-protein interactions for THBS1. THBS1 and its first neighbours are displayed in a circular layout grouped by category (green: expressed in ECs; grey: expressed in tumour; red: expressed in both cell types). Only the direct interactions between molecules up-regulated in tumor and ECs are shown. (THBS1: thrombospondin-1, FBN1: fibrillin-1, ECM1: extracellular matrix protein-1, COL1A1: collagen-1a1, COL1BA1: collagen-1ba1, ITGB3: integrin-b3, TGFB1: transforming growth factor B1, ITGB1: integrin-b1, MMP2: matrix metalloproteinase-2, TNFRSF11B: tumour necrosis factor receptor superfamily, member 11b, IGFBP5: insulin-like growth factor binding protein 5, VEGFA: vascular endothelial growth factor 1, TFP1 : transferrin pseudogene 1, BGN : biglycan, DCN : decorin, JAG1 : jagged 1, ITGA4 : integrin-a4, FN1 : finbronectin-1, COL4A1 : collagen-4a1).
Fig 3: PF is able to reverse hypoxia-induced upregulation of VEGFA, HIF-1a, and p-STAT3 in HRCECs. (a, b) HRCECs were treated with PF (0.5, 5, or 15 µM) for 24 h under normoxia, the level of VEGFA in cell supernatant was detected with ELISA kit. The gene expression of VEGFA and HIF-1a in cells was measured with RT-qPCR. (c, d) HRCECs were treated with PF (5 µM) for 24 h under hypoxia, the level of VEGFA in cell supernatant was detected with ELISA kit. The gene expression of VEGFA and HIF-1a in cells was measured with RT-qPCR. (e, f) HRCECs were treated with PF (5 µM) or/and hypoxia for 24 h, the protein p-STAT3, STAT3, VEGFA, and HIF-1a expression in HRCECs was detected with western blot. *P < 0.05, **P < 0.01 compared with the control group; #P < 0.05, ##P < 0.01 compared with the hypoxia group; n = 3.
Fig 4: Schematic of the fabrication of the scaffold, in vivo and in vitro experiments. a Fabrication of the porous bilayered nanofibrous scaffold. b ADSCs isolated from the bilateral inguinal adipose tissues and then seeded to the scaffold to repair the defect of the urethra. c Hypoxia-preconditioned ADSCs grow in macropores and promote angiogenesis. d The mechanism of hypoxia-preconditioned ADSCs secreted more VEGFA to promote the expression of VEGFR2, HIF-1a, and HK2 to upregulating glycolysis
Supplier Page from Abcam for Human VEGF ELISA Kit