vascular endothelial growth factor vegf a  (Bender MedSystems)

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Loss of the endothelial glycocalyx is associated with increased E-selectin mediated adhesion of lung tumour cells to the brain microvascular endothelium

doi: 10.1186/s13046-015-0223-9

Figure Lengend Snippet: Increased levels of E-selectin detected on the surface of human cerebral microvascular endothelial cells in response to lung tumour CM and secreted-proteins. Confluent hCMEC/D3 cells were stimulated for 4 h with A549 and SK-MES-1 CM ( a ) along with CC ( c ), CL ( d ), IGFBP-7 ( e ), VEGF ( f ) and TNF-α ( g ). E-selectin levels from 3 separate experiments, carried out in quadruplets, were statistically analysed (n ≥ 12, mean ± SD, * p ≤ 0.05 and ** p ≤ 0.01, *** p ≤ 0.005 and **** p ≤ 0.0001). Enhanced E-selectin was also detected within 30 min of CM ( a ) and secreted-proteins ( b ) exposure (* p ≤ 0.05 and ** p ≤ 0.01, n = 6)

Article Snippet: Secreted levels of TNF-α, CL, CC, VEGF-A and IGFBP-7 in the A549 and SK-MES-1 CM were measured by ELISA (TNF-α and CC: Bender MedSystems, Vienna, Austria; CL: R & D systems, Abingdon, UK; VEGF-A: PeproTech®, London, UK) and IGFBP-7: in house ELISA previously described in [ ].

Techniques:

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Loss of the endothelial glycocalyx is associated with increased E-selectin mediated adhesion of lung tumour cells to the brain microvascular endothelium

doi: 10.1186/s13046-015-0223-9

Figure Lengend Snippet: Lung tumour CM and secreted-proteins enhance adhesion of A549 and SK-MES-1 cells to human brain endothelial monolayers. Confluent hCMEC/D3 cells in μ-slides were exposed to A549 CM or SK-MES-1 CM for 30 min and 24 h prior to perfusion with calcein AM stained A549 ( a ) or SK-MES-1 cells ( c ). A549 and SK-MES-1 adhesion to hCMEC/D3 cells treated with 80 ng/ml CC, 10 ng/ml CL, 200 ng/ml IGFBP-7, 0.2 ng/ml VEGF and 160 pg/ml TNF-α at 4 and 24 h ( b ) and ( d ). Mean values from three separate experiments were statistically analysed. * p ≤ 0.05 versus control levels (100 %). e Representative images of adherent calcein-AM stained A549 [( i ) and ( ii )] and SK-MES-1 [( iii ) and ( iv )] cells to DMEM-BS [( i ) and ( iii )] and CL-treated [( ii ) and ( iv )] brain endothelial monolayers. Scale bar = 75 μm

Article Snippet: Secreted levels of TNF-α, CL, CC, VEGF-A and IGFBP-7 in the A549 and SK-MES-1 CM were measured by ELISA (TNF-α and CC: Bender MedSystems, Vienna, Austria; CL: R & D systems, Abingdon, UK; VEGF-A: PeproTech®, London, UK) and IGFBP-7: in house ELISA previously described in [ ].

Techniques: Staining

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Loss of the endothelial glycocalyx is associated with increased E-selectin mediated adhesion of lung tumour cells to the brain microvascular endothelium

doi: 10.1186/s13046-015-0223-9

Figure Lengend Snippet: Exposure of lung tumour CM and secreted-proteins significantly altered the brain endothelial glycocalyx. a and b Integrity of the hCMEC/D3 glycocalyx was assessed by a CBF assay following exposure of ECs for 30 min or 24 h to A549 and SKMES-1 CM ( a ), or 80 ng/ml CC, 10 ng/ml CL, 200 ng/ml IGFBP-7, 0.2 ng/ml VEGF and 160 pg/ml TNF-α ( b ). Data from 6 independent experiments, carried out in sextuplicate, is presented as WGA-FITC fluorescence/μg protein and these values were calculated as a percentage of control. * p ≤ 0.05, ** p ≤ 0.01 vs control level. c Hyaluronan (HA) and syndecan-1 levels were measured by ELISA in hCMEC/D3 growth medium following 30 min treatment with fresh DMEM-BS (control), A549 or SK-MES-1 CM. SK-MES-1 CM vs control = p = 0.007, n = 6. nd = not detected. d and c Representative confocal images of FITC-WGA stained hCMEC/D3 cells before ( d ) and ( e ) after 30 min exposure to SKMES-1 CM. Scale bar = 40 μM

Article Snippet: Secreted levels of TNF-α, CL, CC, VEGF-A and IGFBP-7 in the A549 and SK-MES-1 CM were measured by ELISA (TNF-α and CC: Bender MedSystems, Vienna, Austria; CL: R & D systems, Abingdon, UK; VEGF-A: PeproTech®, London, UK) and IGFBP-7: in house ELISA previously described in [ ].

Techniques: Whole Genome Amplification, Fluorescence, Enzyme-linked Immunosorbent Assay, Staining

Journal: Molecular Vision

Article Title: Vascular endothelial growth factor-C secretion is increased by advanced glycation end-products: possible implication in ocular neovascularization

doi:

Figure Lengend Snippet: Reverse transcription–polymerase chain reaction (RT–PCR) analysis of VEGF-A and VEGF-C. RT–PCR analysis of VEGF-A and VEGF-C mRNA expression, respectively, in ARPE-19 A and HECV B cells cultured 24 h in standard conditions (CTR) or in the presence of AGEs (GS). GAPDH was used as the internal control. VEGF-A and VEGF-C secretion in ARPE-19 C , E: and HECV D , F: cells, respectively, after 24 h culture in CTR or in presence of AGEs (GS). The conditioned media were collected, and ELISA was performed. Concentrations of VEGFs were calculated from standards curves and normalized to total protein. The results are representative of at least three experiments (mean±SEM). *p

Article Snippet: VEGF-A and VEGF-C secretions were assessed with enzyme-linked immunosorbent assay (ELISA; Bender MedSystem, Vienna, Austria).

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Molecular Vision

Article Title: Vascular endothelial growth factor-C secretion is increased by advanced glycation end-products: possible implication in ocular neovascularization

doi:

Figure Lengend Snippet: Secretion of vascular endothelial growth factor (VEGF)-A and –C. VEGF-A and VEGF-C secretion in ARPE-19 ( A , C and HECV B , D cells, respectively, transfected with a scrambled siRNA sequence (siControl) or siRNA of RAGE (siRAGE) following 24 h of culture in standard conditions (CTR) or in the presence of AGEs (GS).The conditioned media were collected, and ELISA was performed. Concentrations of VEGFs were calculated from standards curves and normalized to total protein. The results are representative of at least three experiments (mean±SEM). **p

Article Snippet: VEGF-A and VEGF-C secretions were assessed with enzyme-linked immunosorbent assay (ELISA; Bender MedSystem, Vienna, Austria).

Techniques: Transfection, Sequencing, Enzyme-linked Immunosorbent Assay

Journal: Journal of Clinical Investigation

Article Title: Pathogenesis of persistent lymphatic vessel hyperplasia in chronic airway inflammation

doi: 10.1172/JCI200522037

Figure Lengend Snippet: VEGF-C and VEGF-D in M. pulmonis –infected airways. Immunohistochemical staining of VEGF-C ( A – C ) and VEGF-D ( D – F ) in mouse airways and lung 14 days after infection. ( A ) VEGF-C (green) in epithelium, peribronchial inflammatory cells,

Article Snippet: VEGF-C and VEGF-D in the lungs of nude mice treated intranasally with adenoviruses encoding LacZ, VEGF165, VEGF-C, or VEGF-D ΔNΔC 10 days previously were measured by ELISA (human VEGF-C ELISA [no. BMS297; Bender MedSystems], human VEGF-D ELISA [no. DVED00; R & D Systems]).

Techniques: Infection, Immunohistochemistry, Staining

Journal: BMC Cancer

Article Title: Overexpression of vascular endothelial growth factor C increases growth and alters the metastatic pattern of orthotopic PC-3 prostate tumors

doi: 10.1186/1471-2407-9-362

Figure Lengend Snippet: Effect of VEGF-C overexpression on angiogenesis of orthotopic tumors . H E-staining of representative PC-3/VEGF-C (A, orthotopic; C, subcutaneous) tumors and PC-3/mock (B, orthotopic; D, subcutaneous) tumors. PC-3/VEGF-C tumors showed angiogenic morphology with a rich network of capillaries compared with PC-3/mock tumors. There were significantly more blood capillaries (CD34 positive, arrows) in the PC-3/VEGF-C tumors (E, 220 ± 15 μm/mm 2 , n = 29) compared with PC-3/mock tumors (F and G, 37 ± 6 μm/mm 2 , n = 24), p

Article Snippet: VEGF-C protein concentrations from cell lysates and 24 h conditioned media of cell clones were further quantified by ELISA (Bender MedSystems, Vienna, Austria) according to the manufacturer's instructions.

Techniques: Over Expression, Staining

Journal: BMC Cancer

Article Title: Overexpression of vascular endothelial growth factor C increases growth and alters the metastatic pattern of orthotopic PC-3 prostate tumors

doi: 10.1186/1471-2407-9-362

Figure Lengend Snippet: Effect of VEGFR3-Ig treatment on orthotopic PC-3/VEGF-C tumors . VEGFR3-Ig fusion protein-producing construct or lacZ-Ig controls were injected intravenously into mice five minutes before orthotopic inoculation of PC-3/VEGF-C cells. The density of m-LYVE-1 positive lymphatic capillaries was decreased in VEGFR3-Ig-treated tumors (23 ± 4 μm/mm 2 , n = 8 vs . 123 ± 7 μm/mm 2 , n = 6, p

Article Snippet: VEGF-C protein concentrations from cell lysates and 24 h conditioned media of cell clones were further quantified by ELISA (Bender MedSystems, Vienna, Austria) according to the manufacturer's instructions.

Techniques: Construct, Injection, Mouse Assay

Journal: BMC Cancer

Article Title: Overexpression of vascular endothelial growth factor C increases growth and alters the metastatic pattern of orthotopic PC-3 prostate tumors

doi: 10.1186/1471-2407-9-362

Figure Lengend Snippet: Ectopic expression of VEGF-C in PC-3 cells and orthotopic tumors . PC-3 cells were stably transfected with a vector containing human VEGF-C (PC-3/VEGF-C) or empty vector (mock). One cell clone expressing VEGF-C at a high level, and one control clone (PC-3/mock) were selected for further studies. ELISA analysis of VEGF-C protein concentration in cell lysates (A) and in conditioned media (B) revealed increased expression in PC-3/VEGF-C cells compared with PC-3/mock cells. Data are expressed as means ± SD ng/mL, corresponding 10 6 cells. (C) Western blot analysis revealed that PC-3 cell clones expressed mainly 29/31 kD form of VEGF-C. Detectable levels of 21 kD form were not seen. β-actin control was from cell lysates corresponding to the collected conditioned medium.

Article Snippet: VEGF-C protein concentrations from cell lysates and 24 h conditioned media of cell clones were further quantified by ELISA (Bender MedSystems, Vienna, Austria) according to the manufacturer's instructions.

Techniques: Expressing, Stable Transfection, Transfection, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Protein Concentration, Western Blot, Clone Assay

Journal: BMC Cancer

Article Title: Overexpression of vascular endothelial growth factor C increases growth and alters the metastatic pattern of orthotopic PC-3 prostate tumors

doi: 10.1186/1471-2407-9-362

Figure Lengend Snippet: Effect of VEGF-C overexpression on PC-3 prostate tumor growth . The occurrence of subcutaneous tumors of both PC-3/VEGF-C ( n = 8) and PC-3/mock ( n = 12) was 100%. (A) Subcutaneous PC-3/VEGF-C tumors grew larger (1132 ± 34 mm 3 , gray bars) than PC-3/mock tumors (47 ± 7 mm 3 , black bars) at 4 weeks and the differences in tumor volumes between the groups were significant at all time points between 2 and 4 weeks ( p

Article Snippet: VEGF-C protein concentrations from cell lysates and 24 h conditioned media of cell clones were further quantified by ELISA (Bender MedSystems, Vienna, Austria) according to the manufacturer's instructions.

Techniques: Over Expression

Journal: BMC Cancer

Article Title: Overexpression of vascular endothelial growth factor C increases growth and alters the metastatic pattern of orthotopic PC-3 prostate tumors

doi: 10.1186/1471-2407-9-362

Figure Lengend Snippet: Effect of ectopic VEGF-C on lymphatic vessel density of orthotopic PC-3 tumors . M-LYVE-1-positive staining was observed primarily in the tumor periphery and in the peritumoral area (A-B). No differences were detected in the density of lymphatic vessels Bar 500 μm.

Article Snippet: VEGF-C protein concentrations from cell lysates and 24 h conditioned media of cell clones were further quantified by ELISA (Bender MedSystems, Vienna, Austria) according to the manufacturer's instructions.

Techniques: Staining

Journal: BMC Cancer

Article Title: Overexpression of vascular endothelial growth factor C increases growth and alters the metastatic pattern of orthotopic PC-3 prostate tumors

doi: 10.1186/1471-2407-9-362

Figure Lengend Snippet: Metastasis of PC-3/VEGF-C tumors to prostate-draining lymph nodes and lungs . The prostate draining lymph nodes and the lungs were cut through to detect metastasis. Altogether 8-10 lymph nodes and six lung sections/mouse were studied and number of metastases was counted. The relative metastatic area of lymph nodes was determined by histomorphometry but there were no statistically significant differences (data not shown). The occurrence of metastasis in lymph nodes was 21% ( n = 6 mice out of 29 tumor-bearing mice) in the PC-3/VEGF-C group and 58% ( n = 14 mice out of 24 tumor-bearing mice) in the PC-3/mock group and in lungs was 48% ( n = 14 out of 29 tumor bearing mice) in the PC-3/VEGF-C group and 8% ( n = 2 out of 24 tumor-bearing mice) in the PC-3/mock group.

Article Snippet: VEGF-C protein concentrations from cell lysates and 24 h conditioned media of cell clones were further quantified by ELISA (Bender MedSystems, Vienna, Austria) according to the manufacturer's instructions.

Techniques: Mouse Assay