Structured Review

Millipore p vegfr 1
Ligand-induced activation <t>VEGFR-1/-3</t> activation and downstream AKT and ERK activation by immunoblotting. A, SW480 cells cultured in serum-free media were pretreated with or without cediranib (100 nmol/L), were left untreated (SF; lanes 1, 6 ), or were
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Images

1) Product Images from "Targeting vascular endothelial growth factor receptor-1 and -3 with cediranib (AZD2171): effects on migration and invasion of gastrointestinal cancer cell lines"

Article Title: Targeting vascular endothelial growth factor receptor-1 and -3 with cediranib (AZD2171): effects on migration and invasion of gastrointestinal cancer cell lines

Journal: Molecular cancer therapeutics

doi: 10.1158/1535-7163.MCT-09-0380

Ligand-induced activation VEGFR-1/-3 activation and downstream AKT and ERK activation by immunoblotting. A, SW480 cells cultured in serum-free media were pretreated with or without cediranib (100 nmol/L), were left untreated (SF; lanes 1, 6 ), or were
Figure Legend Snippet: Ligand-induced activation VEGFR-1/-3 activation and downstream AKT and ERK activation by immunoblotting. A, SW480 cells cultured in serum-free media were pretreated with or without cediranib (100 nmol/L), were left untreated (SF; lanes 1, 6 ), or were

Techniques Used: Activation Assay, Cell Culture

2) Product Images from "The Biflavonoid Amentoflavone Inhibits Neovascularization Preventing the Activity of Proangiogenic Vascular Endothelial Growth Factors *"

Article Title: The Biflavonoid Amentoflavone Inhibits Neovascularization Preventing the Activity of Proangiogenic Vascular Endothelial Growth Factors *

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M110.186239

Inhibitory and binding properties of amentoflavone. A , AF structure. B , dose-dependent inhibition of the PlGF-1/VEGFR-1 ( filled circles ), VEGF-A/VEGFR-1 ( open squares ), and VEGF-A/VEGFR-2 ( open triangles ) interaction exerted by AF at concentration ranging between 3.75 and 200 μ m in ELISA-based assays. As a negative control of inhibition fraction G ( supplemental Fig. S2 ) was used at same concentration for PlGF-1/VEGFR-1 ( filled triangles ) and VEGF-A/VEGFR-2 ( open circles ) interactions. Each point was done in triplicate, and each experiment was repeated twice. Data are represented as the mean ± S.E. C , dose-dependent inhibition of VEGF-B/VEGFR-1 ( filled circles ) and PDGF-B/PDGFR-β ( open squares ) interactions exerted by AF at concentration ranging between 3.75 and 200 μ m in ELISA-based assays. Each point was done in triplicate, and each experiment was repeated twice. Data are represented as the mean ± S.E. D–F , sensograms obtained injecting increasing concentrations (from 0.05 to 10 μ m ) of AF on PlGF-1-, VEGF-A-, and human serum albumin-coated Biacore chips.
Figure Legend Snippet: Inhibitory and binding properties of amentoflavone. A , AF structure. B , dose-dependent inhibition of the PlGF-1/VEGFR-1 ( filled circles ), VEGF-A/VEGFR-1 ( open squares ), and VEGF-A/VEGFR-2 ( open triangles ) interaction exerted by AF at concentration ranging between 3.75 and 200 μ m in ELISA-based assays. As a negative control of inhibition fraction G ( supplemental Fig. S2 ) was used at same concentration for PlGF-1/VEGFR-1 ( filled triangles ) and VEGF-A/VEGFR-2 ( open circles ) interactions. Each point was done in triplicate, and each experiment was repeated twice. Data are represented as the mean ± S.E. C , dose-dependent inhibition of VEGF-B/VEGFR-1 ( filled circles ) and PDGF-B/PDGFR-β ( open squares ) interactions exerted by AF at concentration ranging between 3.75 and 200 μ m in ELISA-based assays. Each point was done in triplicate, and each experiment was repeated twice. Data are represented as the mean ± S.E. D–F , sensograms obtained injecting increasing concentrations (from 0.05 to 10 μ m ) of AF on PlGF-1-, VEGF-A-, and human serum albumin-coated Biacore chips.

Techniques Used: Binding Assay, Inhibition, Concentration Assay, Enzyme-linked Immunosorbent Assay, Negative Control

Amentoflavone inhibits VEGFR phosphorylation and HUVEC migration. A , representative pictures of Western blot analysis of VEGFR-1 phosphorylation (P-VEGFR-1) induced by 20 ng/ml of PlGF-1 or 50 ng/ml VEGF-A on cells overexpressing VEGFR-1 (293-Flt-1). AF was used at a concentration ranging between 2 and 25 μ m . As control of inhibition neutralizing antibody anti-PlGF (α-PlGF) or anti-VEGF-A (α-VEGF-A) were used at 3.3 n m . Anti-VEGFR-1 antibody was used for normalization. B , Western blot analysis of VEGFR-2 phosphorylation (P-VEGFR-2) performed in the same condition as reported in A but on HUVECs. Anti-VEGFR-2 antibody was used for normalization. Histograms represent densitometry analysis of three independent experiments, and data are represented as the mean ± S.E. ADU , arbitrary densitometric unit. *, p
Figure Legend Snippet: Amentoflavone inhibits VEGFR phosphorylation and HUVEC migration. A , representative pictures of Western blot analysis of VEGFR-1 phosphorylation (P-VEGFR-1) induced by 20 ng/ml of PlGF-1 or 50 ng/ml VEGF-A on cells overexpressing VEGFR-1 (293-Flt-1). AF was used at a concentration ranging between 2 and 25 μ m . As control of inhibition neutralizing antibody anti-PlGF (α-PlGF) or anti-VEGF-A (α-VEGF-A) were used at 3.3 n m . Anti-VEGFR-1 antibody was used for normalization. B , Western blot analysis of VEGFR-2 phosphorylation (P-VEGFR-2) performed in the same condition as reported in A but on HUVECs. Anti-VEGFR-2 antibody was used for normalization. Histograms represent densitometry analysis of three independent experiments, and data are represented as the mean ± S.E. ADU , arbitrary densitometric unit. *, p

Techniques Used: Migration, Western Blot, Concentration Assay, Inhibition

Docking analysis of PlGF-1·amentoflavone complex. Shown is a surface plot of PlGF-1 showing the residues involved in Flt-1 recognition in green ( A ), the peptide-(94–110), identified by UV cross-linking analysis, and the residues Gln-27 and Arg-32, evidenced with limited proteolysis analysis, highlighted in blue and yellow , respectively ( B ). C , surface plot of the model of the PlGF-1·AF complex showing the lowest binding energy. Red indicates the area within 6 Å from the ligand. A high overlapping with blue and green residues was observed. D , magnification of interaction area between AF and PlGF-1. The PlGF-1 residues of the two monomers ( green and cyan ) involved in AF interaction are indicated. AF and side chains of PlGF-1 residues involved in contacts are represented as sticks with colored atoms ( white , hydrogen; red , oxygen; blue , nitrogen). E , the PlGF-1·VEGFR-1 D2 complex (Protein Data Bank code 1RV6 ) is shown with the two monomers of PlGF-1 homodimer represented in green and cyan , and the VEGFR-1 D2 domain is represented in yellow , whereas the PlGF-1 residues and corresponding side chains at 6 Å from AF inhibitor are highlighted in red .
Figure Legend Snippet: Docking analysis of PlGF-1·amentoflavone complex. Shown is a surface plot of PlGF-1 showing the residues involved in Flt-1 recognition in green ( A ), the peptide-(94–110), identified by UV cross-linking analysis, and the residues Gln-27 and Arg-32, evidenced with limited proteolysis analysis, highlighted in blue and yellow , respectively ( B ). C , surface plot of the model of the PlGF-1·AF complex showing the lowest binding energy. Red indicates the area within 6 Å from the ligand. A high overlapping with blue and green residues was observed. D , magnification of interaction area between AF and PlGF-1. The PlGF-1 residues of the two monomers ( green and cyan ) involved in AF interaction are indicated. AF and side chains of PlGF-1 residues involved in contacts are represented as sticks with colored atoms ( white , hydrogen; red , oxygen; blue , nitrogen). E , the PlGF-1·VEGFR-1 D2 complex (Protein Data Bank code 1RV6 ) is shown with the two monomers of PlGF-1 homodimer represented in green and cyan , and the VEGFR-1 D2 domain is represented in yellow , whereas the PlGF-1 residues and corresponding side chains at 6 Å from AF inhibitor are highlighted in red .

Techniques Used: Binding Assay

3) Product Images from "Calcium dobesilate reduces VEGF signaling by interfering with heparan sulfate binding site and protects from vascular complications in diabetic mice"

Article Title: Calcium dobesilate reduces VEGF signaling by interfering with heparan sulfate binding site and protects from vascular complications in diabetic mice

Journal: PLoS ONE

doi: 10.1371/journal.pone.0218494

Inhibition of endothelial cell proliferation, invasion and migration by CaD. HUVECs were plated in 96-well plates, allowed to attach overnight, and then cultured for 24 h-48 h with CaD in the presence of VEGF 165 (25 ng/ml)-CaD mixture (A) at the indicated concentration. The proliferation was measured as described in the Materials and Methods section. (B) Serum-starved HUVECs were allowed to migrate through trans-well membranes towards a vehicle, VEGF 165 (25 ng/ml) and/or CaD at the indicated concentration for 24 h. Cells that had migrated to the underside of the membrane were processed for calcein-AM staining as described in the materials and methods section. (C) A monolayer of HUVECs was scratched and fresh medium containing vehicle, VEGF and/or CaD was then added. After 14 h, migration distance of HUVECs was quantified. Original magnification, 40x. (D) HUVECs were incubated with vehicle, VEGF 165 (25 ng/ml) and CaD for 15 min. Cells were stained with phalloidin-Alexa Fluor 488 (left images) and DAPI (middle images). Merged view is represented in the right images. Original magnification, 40x. Scale bars represent 5 μm. The results shown are the means ± SD of four independent experiments conducted in triplicate. *P
Figure Legend Snippet: Inhibition of endothelial cell proliferation, invasion and migration by CaD. HUVECs were plated in 96-well plates, allowed to attach overnight, and then cultured for 24 h-48 h with CaD in the presence of VEGF 165 (25 ng/ml)-CaD mixture (A) at the indicated concentration. The proliferation was measured as described in the Materials and Methods section. (B) Serum-starved HUVECs were allowed to migrate through trans-well membranes towards a vehicle, VEGF 165 (25 ng/ml) and/or CaD at the indicated concentration for 24 h. Cells that had migrated to the underside of the membrane were processed for calcein-AM staining as described in the materials and methods section. (C) A monolayer of HUVECs was scratched and fresh medium containing vehicle, VEGF and/or CaD was then added. After 14 h, migration distance of HUVECs was quantified. Original magnification, 40x. (D) HUVECs were incubated with vehicle, VEGF 165 (25 ng/ml) and CaD for 15 min. Cells were stained with phalloidin-Alexa Fluor 488 (left images) and DAPI (middle images). Merged view is represented in the right images. Original magnification, 40x. Scale bars represent 5 μm. The results shown are the means ± SD of four independent experiments conducted in triplicate. *P

Techniques Used: Inhibition, Migration, Cell Culture, Concentration Assay, Staining, Incubation

4) Product Images from "Obesity but not high-fat diet impairs lymphatic function"

Article Title: Obesity but not high-fat diet impairs lymphatic function

Journal: International Journal of Obesity (2005)

doi: 10.1038/ijo.2016.96

Obesity downregulates expression of lymphatic markers in LECs. ( a ) Quantification of mRNA expression in LECs sorted from NCD- and HFD-fed mice skin tissues for all groups, demonstrating relative expression of Prox1, VEGFR-3, CCL21, ICAM-1 and Bax ( n =4–6/group). Data are presented as fold change relative to the NCD-fed controls in all groups. Note the downregulation of Prox1, VEGFR-3 and CCL21 (* P
Figure Legend Snippet: Obesity downregulates expression of lymphatic markers in LECs. ( a ) Quantification of mRNA expression in LECs sorted from NCD- and HFD-fed mice skin tissues for all groups, demonstrating relative expression of Prox1, VEGFR-3, CCL21, ICAM-1 and Bax ( n =4–6/group). Data are presented as fold change relative to the NCD-fed controls in all groups. Note the downregulation of Prox1, VEGFR-3 and CCL21 (* P

Techniques Used: Expressing, Mouse Assay

Exposure of LECs to SA in vitro downregulates expression of VEGFR-3 and its downstream mediators. ( a , b ) Representative western blot images ( a ) for VEGFR-3, p-AKT, p-eNOS and quantification ( b ) normalized to GAPDH expression of total cellular protein harvested from LECs treated with 1 μ m SA, 10 μ m SA+0.3 n m PTENi, PTENi (0.3 n m ), SA+100 ng ml −1 VEGF-C, VEGF-C (100 ng ml −1 ), SA+100 n m insulin or insulin (100 n m ) ( n =4–5/group, * P
Figure Legend Snippet: Exposure of LECs to SA in vitro downregulates expression of VEGFR-3 and its downstream mediators. ( a , b ) Representative western blot images ( a ) for VEGFR-3, p-AKT, p-eNOS and quantification ( b ) normalized to GAPDH expression of total cellular protein harvested from LECs treated with 1 μ m SA, 10 μ m SA+0.3 n m PTENi, PTENi (0.3 n m ), SA+100 ng ml −1 VEGF-C, VEGF-C (100 ng ml −1 ), SA+100 n m insulin or insulin (100 n m ) ( n =4–5/group, * P

Techniques Used: In Vitro, Expressing, Western Blot

5) Product Images from "Obesity but not high-fat diet impairs lymphatic function"

Article Title: Obesity but not high-fat diet impairs lymphatic function

Journal: International Journal of Obesity (2005)

doi: 10.1038/ijo.2016.96

Obesity downregulates expression of lymphatic markers in LECs. ( a ) Quantification of mRNA expression in LECs sorted from NCD- and HFD-fed mice skin tissues for all groups, demonstrating relative expression of Prox1, VEGFR-3, CCL21, ICAM-1 and Bax ( n =4–6/group). Data are presented as fold change relative to the NCD-fed controls in all groups. Note the downregulation of Prox1, VEGFR-3 and CCL21 (* P
Figure Legend Snippet: Obesity downregulates expression of lymphatic markers in LECs. ( a ) Quantification of mRNA expression in LECs sorted from NCD- and HFD-fed mice skin tissues for all groups, demonstrating relative expression of Prox1, VEGFR-3, CCL21, ICAM-1 and Bax ( n =4–6/group). Data are presented as fold change relative to the NCD-fed controls in all groups. Note the downregulation of Prox1, VEGFR-3 and CCL21 (* P

Techniques Used: Expressing, Mouse Assay

Exposure of LECs to SA in vitro downregulates expression of VEGFR-3 and its downstream mediators. ( a , b ) Representative western blot images ( a ) for VEGFR-3, p-AKT, p-eNOS and quantification ( b ) normalized to GAPDH expression of total cellular protein harvested from LECs treated with 1 μ m SA, 10 μ m SA+0.3 n m PTENi, PTENi (0.3 n m ), SA+100 ng ml −1 VEGF-C, VEGF-C (100 ng ml −1 ), SA+100 n m insulin or insulin (100 n m ) ( n =4–5/group, * P
Figure Legend Snippet: Exposure of LECs to SA in vitro downregulates expression of VEGFR-3 and its downstream mediators. ( a , b ) Representative western blot images ( a ) for VEGFR-3, p-AKT, p-eNOS and quantification ( b ) normalized to GAPDH expression of total cellular protein harvested from LECs treated with 1 μ m SA, 10 μ m SA+0.3 n m PTENi, PTENi (0.3 n m ), SA+100 ng ml −1 VEGF-C, VEGF-C (100 ng ml −1 ), SA+100 n m insulin or insulin (100 n m ) ( n =4–5/group, * P

Techniques Used: In Vitro, Expressing, Western Blot

6) Product Images from "Novel application assigned to toluquinol: inhibition of lymphangiogenesis by interfering with VEGF‐C/VEGFR‐3 signalling pathway) Novel application assigned to toluquinol: inhibition of lymphangiogenesis by interfering with VEGF‐C/VEGFR‐3 signalling pathway"

Article Title: Novel application assigned to toluquinol: inhibition of lymphangiogenesis by interfering with VEGF‐C/VEGFR‐3 signalling pathway) Novel application assigned to toluquinol: inhibition of lymphangiogenesis by interfering with VEGF‐C/VEGFR‐3 signalling pathway

Journal: British Journal of Pharmacology

doi: 10.1111/bph.13488

Toluquinol suppresses VEGFR‐3 phosphorylation and downstream signalling targets. LECs were cultured in serum‐depleted conditions for 24 h, and incubated or not with different toluquinol concentrations for 2 h and then stimulated for 30
Figure Legend Snippet: Toluquinol suppresses VEGFR‐3 phosphorylation and downstream signalling targets. LECs were cultured in serum‐depleted conditions for 24 h, and incubated or not with different toluquinol concentrations for 2 h and then stimulated for 30

Techniques Used: Cell Culture, Incubation

7) Product Images from "The FAK scaffold inhibitor C4 disrupts FAK-VEGFR-3 signaling and inhibits pancreatic cancer growth"

Article Title: The FAK scaffold inhibitor C4 disrupts FAK-VEGFR-3 signaling and inhibits pancreatic cancer growth

Journal: Oncotarget

doi:

Compound C4 caused dose- and time-dependent decrease of viability of pancreatic cancer cells, dephosphorylation of FAK and VEGFR-3, and decrease of their complex formation
Figure Legend Snippet: Compound C4 caused dose- and time-dependent decrease of viability of pancreatic cancer cells, dephosphorylation of FAK and VEGFR-3, and decrease of their complex formation

Techniques Used: De-Phosphorylation Assay

8) Product Images from "The FAK scaffold inhibitor C4 disrupts FAK-VEGFR-3 signaling and inhibits pancreatic cancer growth"

Article Title: The FAK scaffold inhibitor C4 disrupts FAK-VEGFR-3 signaling and inhibits pancreatic cancer growth

Journal: Oncotarget

doi:

Compound C4 caused dose- and time-dependent decrease of viability of pancreatic cancer cells, dephosphorylation of FAK and VEGFR-3, and decrease of their complex formation
Figure Legend Snippet: Compound C4 caused dose- and time-dependent decrease of viability of pancreatic cancer cells, dephosphorylation of FAK and VEGFR-3, and decrease of their complex formation

Techniques Used: De-Phosphorylation Assay

9) Product Images from "Wnt3a is critical for endothelial progenitor cell-mediated neural stem cell proliferation and differentiation"

Article Title: Wnt3a is critical for endothelial progenitor cell-mediated neural stem cell proliferation and differentiation

Journal: Molecular Medicine Reports

doi: 10.3892/mmr.2016.5582

Isolation and culture of bone marrow-derived EPCs and spinal cord-derived NSCs. Images show bone marrow-derived EPCs (A) 14 days following isolation and (B) following DAPI staining. The (C) expression of vascular endothelial growth factor-2 and (D) CD133, the (E) uptake of DiI-labeled acetylated low-density lipoprotein and (F) binding to fluorescein isothiocyanate UEA-1 were analyzed. (G) Spinal cord-derived NSCs 21 days following isolation, (H) expressing nestin and (I) following DAPI staining. Scale bar=50 µ m. EPCs, endothelial progenitor cells; NSCs, neural stem cells; DAPI, 2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride.
Figure Legend Snippet: Isolation and culture of bone marrow-derived EPCs and spinal cord-derived NSCs. Images show bone marrow-derived EPCs (A) 14 days following isolation and (B) following DAPI staining. The (C) expression of vascular endothelial growth factor-2 and (D) CD133, the (E) uptake of DiI-labeled acetylated low-density lipoprotein and (F) binding to fluorescein isothiocyanate UEA-1 were analyzed. (G) Spinal cord-derived NSCs 21 days following isolation, (H) expressing nestin and (I) following DAPI staining. Scale bar=50 µ m. EPCs, endothelial progenitor cells; NSCs, neural stem cells; DAPI, 2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride.

Techniques Used: Isolation, Derivative Assay, Staining, Expressing, Labeling, Binding Assay

10) Product Images from "Regulation of lymphangiogenesis in the diaphragm by macrophages and VEGFR-3 signaling"

Article Title: Regulation of lymphangiogenesis in the diaphragm by macrophages and VEGFR-3 signaling

Journal: Angiogenesis

doi: 10.1007/s10456-016-9523-8

Colony-stimulating factor-1 receptor-positive macrophages negatively regulate lymphatic vessel branching on the pleural side of the diaphragm. P6 diaphragm whole mounts show CSF-1R and LYVE-1 double-positive macrophages in proximity to LYVE-1-positive lymphatic vessels ( a ). Compared to WT littermates ( b ), CD206 ( cyan )- and CD68 ( red )-stained whole mounts of diaphragms of P7 Csf1r −/− pups show depletion of macrophages ( c ). Segments of pleural diaphragm LYVE-1 whole mounts of P7 WT ( d ) or Csf1r −/− pups ( e ). CD68-stained whole mounts of IgG ( f )- or AFS98 ( g )-treated pups show depletion of macrophages on the pleural side of the diaphragmatic muscle by AFS98 treatment. Quantification of the CD68-positive area per visual field in the diaphragm ( h ). Dots represent mean values per mouse, and lines indicate the group means. Quantification of diaphragmatic lymphatic vessel development of Csf1r −/− pups shows a significant increase in branches and lymphatic loops compared to the WT controls ( k – o ). Dots represent mean values per mouse, and lines indicate the group means. Segments of pleural diaphragm LYVE-1 whole mounts of P7 IgG ( i )- or AFS98-treated pups ( j ). Quantification of diaphragmatic lymphatic vessel development of P7 AFS98-treated pups shows a significant increase in branches, lymphatic loops and a significant decrease in average branch length compared to the IgG controls ( p – t ). Dots represent mean values per mouse, and lines indicate the group means. Quantifications of LEC sprouts per bead show that isolated P7 diaphragmatic macrophage-conditioned medium significantly decreased VEGF-A/FGF-induced LEC sprouting ( u ). Dots represent mean values per well, and lines indicate the group means. Scale bars a 50 µm, b and c 50 µm, d and e 1 mm, f and g 50 µm, i and j 1 mm. * p
Figure Legend Snippet: Colony-stimulating factor-1 receptor-positive macrophages negatively regulate lymphatic vessel branching on the pleural side of the diaphragm. P6 diaphragm whole mounts show CSF-1R and LYVE-1 double-positive macrophages in proximity to LYVE-1-positive lymphatic vessels ( a ). Compared to WT littermates ( b ), CD206 ( cyan )- and CD68 ( red )-stained whole mounts of diaphragms of P7 Csf1r −/− pups show depletion of macrophages ( c ). Segments of pleural diaphragm LYVE-1 whole mounts of P7 WT ( d ) or Csf1r −/− pups ( e ). CD68-stained whole mounts of IgG ( f )- or AFS98 ( g )-treated pups show depletion of macrophages on the pleural side of the diaphragmatic muscle by AFS98 treatment. Quantification of the CD68-positive area per visual field in the diaphragm ( h ). Dots represent mean values per mouse, and lines indicate the group means. Quantification of diaphragmatic lymphatic vessel development of Csf1r −/− pups shows a significant increase in branches and lymphatic loops compared to the WT controls ( k – o ). Dots represent mean values per mouse, and lines indicate the group means. Segments of pleural diaphragm LYVE-1 whole mounts of P7 IgG ( i )- or AFS98-treated pups ( j ). Quantification of diaphragmatic lymphatic vessel development of P7 AFS98-treated pups shows a significant increase in branches, lymphatic loops and a significant decrease in average branch length compared to the IgG controls ( p – t ). Dots represent mean values per mouse, and lines indicate the group means. Quantifications of LEC sprouts per bead show that isolated P7 diaphragmatic macrophage-conditioned medium significantly decreased VEGF-A/FGF-induced LEC sprouting ( u ). Dots represent mean values per well, and lines indicate the group means. Scale bars a 50 µm, b and c 50 µm, d and e 1 mm, f and g 50 µm, i and j 1 mm. * p

Techniques Used: Staining, Isolation

11) Product Images from "Novel Vascular Endothelial Growth Factor D Variants with Increased Biological Activity *"

Article Title: Novel Vascular Endothelial Growth Factor D Variants with Increased Biological Activity *

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M109.001123

Screening of the biological activity of the VEGF-D mutants on Ba/F3-VEGFR-2 and Ba/F3-VEGFR-3 cell lines. VEGF-D ΔNΔC and the mutants were produced in transiently transfected 293T cells. The concentrations of VEGF-D ΔNΔC
Figure Legend Snippet: Screening of the biological activity of the VEGF-D mutants on Ba/F3-VEGFR-2 and Ba/F3-VEGFR-3 cell lines. VEGF-D ΔNΔC and the mutants were produced in transiently transfected 293T cells. The concentrations of VEGF-D ΔNΔC

Techniques Used: Activity Assay, Produced, Transfection

Phosphorylation of VEGFR-2, VEGFR-3, and Akt. Serum-starved cells were incubated with recombinant VEGF proteins, and the phosphorylation status of VEGFRs or Akt was analyzed from cell lysates. The results are representatives of replicate experiments yielding
Figure Legend Snippet: Phosphorylation of VEGFR-2, VEGFR-3, and Akt. Serum-starved cells were incubated with recombinant VEGF proteins, and the phosphorylation status of VEGFRs or Akt was analyzed from cell lysates. The results are representatives of replicate experiments yielding

Techniques Used: Incubation, Recombinant

Determination of biological activity and receptor binding affinity of the purified recombinant VEGFs. Ba/F3-VEGFR-2 ( A ) and Ba/F3-VEGFR-3 ( B ) cell growth and survival assays were performed with purified recombinant VEGF proteins. C , the relative VEGFR-2
Figure Legend Snippet: Determination of biological activity and receptor binding affinity of the purified recombinant VEGFs. Ba/F3-VEGFR-2 ( A ) and Ba/F3-VEGFR-3 ( B ) cell growth and survival assays were performed with purified recombinant VEGF proteins. C , the relative VEGFR-2

Techniques Used: Activity Assay, Binding Assay, Purification, Recombinant

12) Product Images from "Calcium dobesilate reduces VEGF signaling by interfering with heparan sulfate binding site and protects from vascular complications in diabetic mice"

Article Title: Calcium dobesilate reduces VEGF signaling by interfering with heparan sulfate binding site and protects from vascular complications in diabetic mice

Journal: PLoS ONE

doi: 10.1371/journal.pone.0218494

Calcium dobesilate (CaD) inhibits VEGF-induced VEGFR-2 activation. VEGF 165 (25 ng/mL) was premixed and incubated with: ( A ) various CaD concentrations (6–100 μM) or ( B ) 50–100 μM CaD for 1 h, before exposure to HUVECs for 2 min. ( C ) HUVECs were incubated with 50 and 100μM CaD for 60 min with three subsequent washing steps with warm medium before stimulation with VEGF 165 for 2 min. Western blot analysis was performed using anti-phospho-VEGFR-2 antibody and total VEGFR2 was used as a loading control after membrane stripping. Each bar represents the mean ± SD (n = 3). *P
Figure Legend Snippet: Calcium dobesilate (CaD) inhibits VEGF-induced VEGFR-2 activation. VEGF 165 (25 ng/mL) was premixed and incubated with: ( A ) various CaD concentrations (6–100 μM) or ( B ) 50–100 μM CaD for 1 h, before exposure to HUVECs for 2 min. ( C ) HUVECs were incubated with 50 and 100μM CaD for 60 min with three subsequent washing steps with warm medium before stimulation with VEGF 165 for 2 min. Western blot analysis was performed using anti-phospho-VEGFR-2 antibody and total VEGFR2 was used as a loading control after membrane stripping. Each bar represents the mean ± SD (n = 3). *P

Techniques Used: Activation Assay, Incubation, Western Blot, Stripping Membranes

Calcium dobesilate (CaD) inhibits VEGF-induced MEK/ERK MAP kinase activation. CaD at 50, 100 and 200μM was incubated with VEGF (25 ng/mL) for 60 min before exposure to HUVECs for 15 min. Phosphorylated ERK and MEK1/2 was determined by Western blot as described in Figure 2. Data are expressed as mean ± SD (n = 3). * p
Figure Legend Snippet: Calcium dobesilate (CaD) inhibits VEGF-induced MEK/ERK MAP kinase activation. CaD at 50, 100 and 200μM was incubated with VEGF (25 ng/mL) for 60 min before exposure to HUVECs for 15 min. Phosphorylated ERK and MEK1/2 was determined by Western blot as described in Figure 2. Data are expressed as mean ± SD (n = 3). * p

Techniques Used: Activation Assay, Incubation, Western Blot

13) Product Images from "Fluid Pressure Is a Magnitude-Dependent Modulator of Early Endothelial Tubulogenic Activity: Implications Related to a Potential Tissue-Engineering Control Parameter"

Article Title: Fluid Pressure Is a Magnitude-Dependent Modulator of Early Endothelial Tubulogenic Activity: Implications Related to a Potential Tissue-Engineering Control Parameter

Journal: Tissue Engineering. Part A

doi: 10.1089/ten.tea.2011.0588

Pressure upregulates cellular levels of VEGF-C and membrane expression of its high-affinity receptor, VEGFR-3. (A, B) Histograms of fluorescence intensity, that is, for either VEGF-C (A) or VEGFR-3 (B) , were plotted for unlabeled BAEC (no-stain; filled
Figure Legend Snippet: Pressure upregulates cellular levels of VEGF-C and membrane expression of its high-affinity receptor, VEGFR-3. (A, B) Histograms of fluorescence intensity, that is, for either VEGF-C (A) or VEGFR-3 (B) , were plotted for unlabeled BAEC (no-stain; filled

Techniques Used: Expressing, Fluorescence, Staining

14) Product Images from "Rapid vascular regrowth in tumors after reversal of VEGF inhibition"

Article Title: Rapid vascular regrowth in tumors after reversal of VEGF inhibition

Journal: Journal of Clinical Investigation

doi: 10.1172/JCI24612

Selective inhibition of VEGFR signaling by AG-028262 causes reversible changes in endothelial cells and pericytes in RIP-Tag2 tumors.
Figure Legend Snippet: Selective inhibition of VEGFR signaling by AG-028262 causes reversible changes in endothelial cells and pericytes in RIP-Tag2 tumors.

Techniques Used: Inhibition

Changes in pericyte phenotype during regression and regrowth of blood vessels in RIP-Tag2 tumors.
Figure Legend Snippet: Changes in pericyte phenotype during regression and regrowth of blood vessels in RIP-Tag2 tumors.

Techniques Used:

15) Product Images from "Novel Vascular Endothelial Growth Factor D Variants with Increased Biological Activity *"

Article Title: Novel Vascular Endothelial Growth Factor D Variants with Increased Biological Activity *

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M109.001123

Expression, sVEGFR2-Fc binding and dimerization of VEGF-D ΔNΔC variants . A , an immunoblot of transiently transfected 293T cell media separated with SDS-PAGE under reducing conditions using an anti-VEGF-D mAb. B , precipitation of receptor
Figure Legend Snippet: Expression, sVEGFR2-Fc binding and dimerization of VEGF-D ΔNΔC variants . A , an immunoblot of transiently transfected 293T cell media separated with SDS-PAGE under reducing conditions using an anti-VEGF-D mAb. B , precipitation of receptor

Techniques Used: Expressing, Binding Assay, Transfection, SDS Page

16) Product Images from "Transcription factor Dlx2 protects from TGF?-induced cell-cycle arrest and apoptosis"

Article Title: Transcription factor Dlx2 protects from TGF?-induced cell-cycle arrest and apoptosis

Journal: The EMBO Journal

doi: 10.1038/emboj.2011.319

Dlx2 expression attenuates Smad-dependent, canonical TGFβ signalling. ( A ) Immunoblotting analysis of TGFβRII and Smad4 protein levels in Dlx2-expressing and control NMuMG cells is shown. Immunoblotting against vinculin was used as loading
Figure Legend Snippet: Dlx2 expression attenuates Smad-dependent, canonical TGFβ signalling. ( A ) Immunoblotting analysis of TGFβRII and Smad4 protein levels in Dlx2-expressing and control NMuMG cells is shown. Immunoblotting against vinculin was used as loading

Techniques Used: Expressing

17) Product Images from "Regulation of lymphangiogenesis in the diaphragm by macrophages and VEGFR-3 signaling"

Article Title: Regulation of lymphangiogenesis in the diaphragm by macrophages and VEGFR-3 signaling

Journal: Angiogenesis

doi: 10.1007/s10456-016-9523-8

Diaphragmatic lymphatic vessel growth is VEGFR-3 dependent. Segments of diaphragm whole mounts stained for LYVE-1 of P7 WT ( a ) and K14-VEGFR-3-Fc transgenic (TG) littermates ( b ). Quantification of diaphragmatic lymphatic vessel (LV) development of WT and K14-VEGFR-3-Fc transgenic (TG) pups ( c – f ). Dots represent mean values per mouse, and lines indicate the group means. As only 2 out of 6 diaphragm segments of TG pups had detectable vessels, no statistical analysis was performed for the parameters average LV branch length ( e ) and LV diameter ( f ). Segments of LYVE-1-stained pleural diaphragm whole mounts of P5 pups treated with IgG control ( g ) or antibodies blocking VEGFR-1 (mF1) ( h ), VEGFR-2 (DC101) ( i ) or VEGFR-3 (mF4) ( j ) in utero at E16.5 and E18.5. Quantification of diaphragmatic lymphatic vessel development of VEGFR-blocking antibody-treated pups ( k – n ). Dots represent mean values per mouse, and lines indicate the group means. Scale bars 1 mm. * p
Figure Legend Snippet: Diaphragmatic lymphatic vessel growth is VEGFR-3 dependent. Segments of diaphragm whole mounts stained for LYVE-1 of P7 WT ( a ) and K14-VEGFR-3-Fc transgenic (TG) littermates ( b ). Quantification of diaphragmatic lymphatic vessel (LV) development of WT and K14-VEGFR-3-Fc transgenic (TG) pups ( c – f ). Dots represent mean values per mouse, and lines indicate the group means. As only 2 out of 6 diaphragm segments of TG pups had detectable vessels, no statistical analysis was performed for the parameters average LV branch length ( e ) and LV diameter ( f ). Segments of LYVE-1-stained pleural diaphragm whole mounts of P5 pups treated with IgG control ( g ) or antibodies blocking VEGFR-1 (mF1) ( h ), VEGFR-2 (DC101) ( i ) or VEGFR-3 (mF4) ( j ) in utero at E16.5 and E18.5. Quantification of diaphragmatic lymphatic vessel development of VEGFR-blocking antibody-treated pups ( k – n ). Dots represent mean values per mouse, and lines indicate the group means. Scale bars 1 mm. * p

Techniques Used: Staining, Transgenic Assay, Blocking Assay, In Utero

VEGFR-3 blockade leads to similar lymphatic vessel phenotypes in the tail dermis and on the pleural diaphragm side. LYVE-1 whole mounts of tail dermis of P7 WT ( a ) and K14-VEGFR-3-Fc transgenic littermates ( b ). LYVE-1 whole mounts of tail dermis of P5 pups treated with IgG ( c ) or blocking antibodies against VEGFR-1 (mF1) ( d ), VEGFR-2 (DC101) ( e ), or VEGFR-3 (mF4) ( f ) in utero at E16.5 and E18.5. VEGFR-3 blockade leads in both approaches to an almost complete inhibition of the lymphangiogenic process, whereas VEGFR-1 and -2 blockades have no obvious effect. Scale bar 100 µm
Figure Legend Snippet: VEGFR-3 blockade leads to similar lymphatic vessel phenotypes in the tail dermis and on the pleural diaphragm side. LYVE-1 whole mounts of tail dermis of P7 WT ( a ) and K14-VEGFR-3-Fc transgenic littermates ( b ). LYVE-1 whole mounts of tail dermis of P5 pups treated with IgG ( c ) or blocking antibodies against VEGFR-1 (mF1) ( d ), VEGFR-2 (DC101) ( e ), or VEGFR-3 (mF4) ( f ) in utero at E16.5 and E18.5. VEGFR-3 blockade leads in both approaches to an almost complete inhibition of the lymphangiogenic process, whereas VEGFR-1 and -2 blockades have no obvious effect. Scale bar 100 µm

Techniques Used: Transgenic Assay, Blocking Assay, In Utero, Inhibition

18) Product Images from "Antitumour efficacy of MEK inhibitors in human lung cancer cells and their derivatives with acquired resistance to different tyrosine kinase inhibitors"

Article Title: Antitumour efficacy of MEK inhibitors in human lung cancer cells and their derivatives with acquired resistance to different tyrosine kinase inhibitors

Journal: British Journal of Cancer

doi: 10.1038/bjc.2011.244

Growth inhibitory effects of treatment with selective MEK inhibitors in parental and TKI-R CALU-3 cancer cells. MTT cell proliferation assays were performed in parental lung adenocarcinoma CALU-3 cells (P) and in their TKI-R derivatives (ERL-R, GEF-R, VAN-R and SOR-R), treated for 3 days with the indicated concentrations of each of two selective MEK inhibitors, MSC19363669B ( A ) or selumetinib ( C ). Results represent the average (±s.d.) of three separate experiments, each performed in quadruplicate. Western blotting analysis of MEK and MAPK activation following treatment with the indicated concentration of each of two selective MEK inhibitors, MSC19363669B ( B ) or selumetinib ( D ) on TKI-R CALU-3 and on parental cell line ( E ). β -Actin was included as a loading control.
Figure Legend Snippet: Growth inhibitory effects of treatment with selective MEK inhibitors in parental and TKI-R CALU-3 cancer cells. MTT cell proliferation assays were performed in parental lung adenocarcinoma CALU-3 cells (P) and in their TKI-R derivatives (ERL-R, GEF-R, VAN-R and SOR-R), treated for 3 days with the indicated concentrations of each of two selective MEK inhibitors, MSC19363669B ( A ) or selumetinib ( C ). Results represent the average (±s.d.) of three separate experiments, each performed in quadruplicate. Western blotting analysis of MEK and MAPK activation following treatment with the indicated concentration of each of two selective MEK inhibitors, MSC19363669B ( B ) or selumetinib ( D ) on TKI-R CALU-3 and on parental cell line ( E ). β -Actin was included as a loading control.

Techniques Used: MTT Assay, Western Blot, Activation Assay, Concentration Assay

Effects of treatment with selective MEK inhibitors on the invasive, migratory and anchorage-independent colony-forming capabilities of TKI-R CALU-3 cancer cells. Invasion ( A and D ), migration ( B and E ) and anchorage-independent growth ( C and F ) were evaluated in TKI-R lung adenocarcinoma CALU-3 derivatives (ERL-R, GEF-R, VAN-R and SOR-R) after treatment with the indicated concentrations of MSC19363669B ( A – C ) or selumetinib ( D – F ). The results are the average±s.d. of three independent experiments, each done in triplicate.
Figure Legend Snippet: Effects of treatment with selective MEK inhibitors on the invasive, migratory and anchorage-independent colony-forming capabilities of TKI-R CALU-3 cancer cells. Invasion ( A and D ), migration ( B and E ) and anchorage-independent growth ( C and F ) were evaluated in TKI-R lung adenocarcinoma CALU-3 derivatives (ERL-R, GEF-R, VAN-R and SOR-R) after treatment with the indicated concentrations of MSC19363669B ( A – C ) or selumetinib ( D – F ). The results are the average±s.d. of three independent experiments, each done in triplicate.

Techniques Used: Migration

Antitumour activity of the selective MEK inhibitor MSC19363669B in parental and TKI-R CALU-3 xenografts. ( A ) Parental (P) CALU-3 cancer cells; ( B ) ERL-R CALU-3 cancer cells; ( C ) GEF-R CALU-3 cancer cells; ( D ) VAN-R CALU-3 cancer cells; ( E ) SOR-R CALU-3 cancer cells. Athymic nude mice were injected subcutaneously into the dorsal flank with 10 7 cancer cells. After 7 to 10 days (average tumour size, 75 mm 3 ), mice were treated as indicated in Materials and Methods section for 5 weeks. Each treatment group consisted of eight mice. Data represent the average (±s.d.). Student's t -test was used to compare tumour sizes among different treatment groups at day 35 following the start of treatment. ( A ) P-CALU-3: MSC19363669B vs control (two-sided P
Figure Legend Snippet: Antitumour activity of the selective MEK inhibitor MSC19363669B in parental and TKI-R CALU-3 xenografts. ( A ) Parental (P) CALU-3 cancer cells; ( B ) ERL-R CALU-3 cancer cells; ( C ) GEF-R CALU-3 cancer cells; ( D ) VAN-R CALU-3 cancer cells; ( E ) SOR-R CALU-3 cancer cells. Athymic nude mice were injected subcutaneously into the dorsal flank with 10 7 cancer cells. After 7 to 10 days (average tumour size, 75 mm 3 ), mice were treated as indicated in Materials and Methods section for 5 weeks. Each treatment group consisted of eight mice. Data represent the average (±s.d.). Student's t -test was used to compare tumour sizes among different treatment groups at day 35 following the start of treatment. ( A ) P-CALU-3: MSC19363669B vs control (two-sided P

Techniques Used: Activity Assay, Mouse Assay, Injection

Pro-apoptotic activity of two MEK inhibitors, MSC19363669B ( A ) and selumetinib ( B ), on TKI-R CALU-3. TKI-R CALU-3 cells were treated for the indicated time with two MEK inhibitors, MSC19363669B and selumetinib, at the showed doses. Apoptosis was assessed by a modified TUNEL assay as described in Materials and Methods section.
Figure Legend Snippet: Pro-apoptotic activity of two MEK inhibitors, MSC19363669B ( A ) and selumetinib ( B ), on TKI-R CALU-3. TKI-R CALU-3 cells were treated for the indicated time with two MEK inhibitors, MSC19363669B and selumetinib, at the showed doses. Apoptosis was assessed by a modified TUNEL assay as described in Materials and Methods section.

Techniques Used: Activity Assay, Modification, TUNEL Assay

19) Product Images from "Rapamycin Inhibits Lymphatic Endothelial Cell Tube Formation by Downregulating Vascular Endothelial Growth Factor Receptor 3 Protein Expression 1"

Article Title: Rapamycin Inhibits Lymphatic Endothelial Cell Tube Formation by Downregulating Vascular Endothelial Growth Factor Receptor 3 Protein Expression 1

Journal: Neoplasia (New York, N.Y.)

doi:

Rapamycin inhibits cellular protein expression of VEGFR-3 in LECs. LECs (A), or LECs infected for 24 hours with Ad-GFP and Ad-mTOR-T, respectively (B), were treated with rapamycin (Rapa, 100 ng/ml) for 24 hours, in the presence or absence of IGF-1 (10 ng/ml) or 2% FBS, followed by Western blot analysis with the indicated antibodies. β-Tubulin was used as a loading control.
Figure Legend Snippet: Rapamycin inhibits cellular protein expression of VEGFR-3 in LECs. LECs (A), or LECs infected for 24 hours with Ad-GFP and Ad-mTOR-T, respectively (B), were treated with rapamycin (Rapa, 100 ng/ml) for 24 hours, in the presence or absence of IGF-1 (10 ng/ml) or 2% FBS, followed by Western blot analysis with the indicated antibodies. β-Tubulin was used as a loading control.

Techniques Used: Expressing, Infection, Western Blot

20) Product Images from "Enhanced Matrix Metalloproteinase Activity in the Spontaneously Hypertensive Rat: VEGFR-2 Cleavage, Endothelial Apoptosis, and Capillary Rarefaction"

Article Title: Enhanced Matrix Metalloproteinase Activity in the Spontaneously Hypertensive Rat: VEGFR-2 Cleavage, Endothelial Apoptosis, and Capillary Rarefaction

Journal: Journal of Vascular Research

doi: 10.1159/000281582

a Extracellular domain cleavage of VEGFR-2 by SHR plasma, MMP-7 and MMP-9. VEGFR-2 immunolabel density was measured after acute treatment (for 6 h) of Wistar heart muscle sections with plasma from WKY and SHR, and with purified MMP −7 and MMP-9. There is extensive cleavage of the extracellular domain of VEGFR-2 on naïve endothelial cells (arrows). All receptor cleavage by SHR plasma, MMP-7, and MMP-9 was blocked by addition of doxycycline (11.3 μ M , see with doxycycline group) as well as GM6001 (1 μ M ), EDTA (10 m M ; not shown). Measurements are derived from 4 repeats per plasma/MMP group and 8 arterioles per Wistar cardiac section. * p
Figure Legend Snippet: a Extracellular domain cleavage of VEGFR-2 by SHR plasma, MMP-7 and MMP-9. VEGFR-2 immunolabel density was measured after acute treatment (for 6 h) of Wistar heart muscle sections with plasma from WKY and SHR, and with purified MMP −7 and MMP-9. There is extensive cleavage of the extracellular domain of VEGFR-2 on naïve endothelial cells (arrows). All receptor cleavage by SHR plasma, MMP-7, and MMP-9 was blocked by addition of doxycycline (11.3 μ M , see with doxycycline group) as well as GM6001 (1 μ M ), EDTA (10 m M ; not shown). Measurements are derived from 4 repeats per plasma/MMP group and 8 arterioles per Wistar cardiac section. * p

Techniques Used: Immunolabeling, Purification, Derivative Assay

21) Product Images from "Regulation of lymphangiogenesis in the diaphragm by macrophages and VEGFR-3 signaling"

Article Title: Regulation of lymphangiogenesis in the diaphragm by macrophages and VEGFR-3 signaling

Journal: Angiogenesis

doi: 10.1007/s10456-016-9523-8

Colony-stimulating factor-1 receptor-positive macrophages negatively regulate lymphatic vessel branching on the pleural side of the diaphragm. P6 diaphragm whole mounts show CSF-1R and LYVE-1 double-positive macrophages in proximity to LYVE-1-positive lymphatic vessels ( a ). Compared to WT littermates ( b ), CD206 ( cyan )- and CD68 ( red )-stained whole mounts of diaphragms of P7 Csf1r −/− pups show depletion of macrophages ( c ). Segments of pleural diaphragm LYVE-1 whole mounts of P7 WT ( d ) or Csf1r −/− pups ( e ). CD68-stained whole mounts of IgG ( f )- or AFS98 ( g )-treated pups show depletion of macrophages on the pleural side of the diaphragmatic muscle by AFS98 treatment. Quantification of the CD68-positive area per visual field in the diaphragm ( h ). Dots represent mean values per mouse, and lines indicate the group means. Quantification of diaphragmatic lymphatic vessel development of Csf1r −/− pups shows a significant increase in branches and lymphatic loops compared to the WT controls ( k – o ). Dots represent mean values per mouse, and lines indicate the group means. Segments of pleural diaphragm LYVE-1 whole mounts of P7 IgG ( i )- or AFS98-treated pups ( j ). Quantification of diaphragmatic lymphatic vessel development of P7 AFS98-treated pups shows a significant increase in branches, lymphatic loops and a significant decrease in average branch length compared to the IgG controls ( p – t ). Dots represent mean values per mouse, and lines indicate the group means. Quantifications of LEC sprouts per bead show that isolated P7 diaphragmatic macrophage-conditioned medium significantly decreased VEGF-A/FGF-induced LEC sprouting ( u ). Dots represent mean values per well, and lines indicate the group means. Scale bars a 50 µm, b and c 50 µm, d and e 1 mm, f and g 50 µm, i and j 1 mm. * p
Figure Legend Snippet: Colony-stimulating factor-1 receptor-positive macrophages negatively regulate lymphatic vessel branching on the pleural side of the diaphragm. P6 diaphragm whole mounts show CSF-1R and LYVE-1 double-positive macrophages in proximity to LYVE-1-positive lymphatic vessels ( a ). Compared to WT littermates ( b ), CD206 ( cyan )- and CD68 ( red )-stained whole mounts of diaphragms of P7 Csf1r −/− pups show depletion of macrophages ( c ). Segments of pleural diaphragm LYVE-1 whole mounts of P7 WT ( d ) or Csf1r −/− pups ( e ). CD68-stained whole mounts of IgG ( f )- or AFS98 ( g )-treated pups show depletion of macrophages on the pleural side of the diaphragmatic muscle by AFS98 treatment. Quantification of the CD68-positive area per visual field in the diaphragm ( h ). Dots represent mean values per mouse, and lines indicate the group means. Quantification of diaphragmatic lymphatic vessel development of Csf1r −/− pups shows a significant increase in branches and lymphatic loops compared to the WT controls ( k – o ). Dots represent mean values per mouse, and lines indicate the group means. Segments of pleural diaphragm LYVE-1 whole mounts of P7 IgG ( i )- or AFS98-treated pups ( j ). Quantification of diaphragmatic lymphatic vessel development of P7 AFS98-treated pups shows a significant increase in branches, lymphatic loops and a significant decrease in average branch length compared to the IgG controls ( p – t ). Dots represent mean values per mouse, and lines indicate the group means. Quantifications of LEC sprouts per bead show that isolated P7 diaphragmatic macrophage-conditioned medium significantly decreased VEGF-A/FGF-induced LEC sprouting ( u ). Dots represent mean values per well, and lines indicate the group means. Scale bars a 50 µm, b and c 50 µm, d and e 1 mm, f and g 50 µm, i and j 1 mm. * p

Techniques Used: Staining, Isolation

Macrophage depletion does not impair lymphatic vessel drainage function. Stereomicroscope image showing fluorescent microspheres within mediastinal lymph nodes ( a , arrows ) in exposed thorax 1 h after i.p. injection of microspheres into P8 pup. T thymus, H heart, R rib cage. FACS analysis in the FSC/SSC ( b ) and FITC/FSC ( c ) channels. FACS analysis of green–yellow fluorescent microspheres in PBS in mediastinal lymph nodes of uninjected pups ( d ), and in non-draining inguinal lymph nodes ( e ) and in draining mediastinal lymph nodes ( f ) 1 h after i.p. injection of microspheres into WT P8 pups. Quantification by FACS showed that there was no difference in drainage of fluorescent microspheres to pooled mediastinal lymph nodes 1 h after i.p. injection of microspheres into rat IgG- or AFS98-treated P7 pups ( g )
Figure Legend Snippet: Macrophage depletion does not impair lymphatic vessel drainage function. Stereomicroscope image showing fluorescent microspheres within mediastinal lymph nodes ( a , arrows ) in exposed thorax 1 h after i.p. injection of microspheres into P8 pup. T thymus, H heart, R rib cage. FACS analysis in the FSC/SSC ( b ) and FITC/FSC ( c ) channels. FACS analysis of green–yellow fluorescent microspheres in PBS in mediastinal lymph nodes of uninjected pups ( d ), and in non-draining inguinal lymph nodes ( e ) and in draining mediastinal lymph nodes ( f ) 1 h after i.p. injection of microspheres into WT P8 pups. Quantification by FACS showed that there was no difference in drainage of fluorescent microspheres to pooled mediastinal lymph nodes 1 h after i.p. injection of microspheres into rat IgG- or AFS98-treated P7 pups ( g )

Techniques Used: Injection, FACS

22) Product Images from "Autocrine activity of soluble Flt-1 controls endothelial cell function and angiogenesis"

Article Title: Autocrine activity of soluble Flt-1 controls endothelial cell function and angiogenesis

Journal: Vascular Cell

doi: 10.1186/2045-824X-3-15

sFlt-1 inhibits endothelial cell proliferation and VEGF receptor phosphorylation . (a) Cell proliferation after 48 hours of treatment in HUVEC infected with an adenovirus encoding sFlt-1 (Ad-sFlt1) or β-galactosidase control (Ad-βGal). Western blot demonstrating decreased Erk1/2 phosphorylation in HUVEC infected with Ad-sFlt-1 compared with Ad-β-gal control (insert). (b) Dramatic reduction of sFlt-1 release from HUVEC 24 hours after transfection with sFlt-1 siRNA. (c) Cell proliferation after 48 hours in sFlt-1 siRNA transfected HUVEC. (d) Western blot showing VEGF receptor-2 (VEGFR-2) phosphorylation at tyrosine 951 in HUVEC transfected with sFlt-1, or control siRNA. VEGFR-2 and β-actin were used as a loading control. Data are expressed as mean (± SEM) or representative of three or more independent experiments performed in triplicate. ** P
Figure Legend Snippet: sFlt-1 inhibits endothelial cell proliferation and VEGF receptor phosphorylation . (a) Cell proliferation after 48 hours of treatment in HUVEC infected with an adenovirus encoding sFlt-1 (Ad-sFlt1) or β-galactosidase control (Ad-βGal). Western blot demonstrating decreased Erk1/2 phosphorylation in HUVEC infected with Ad-sFlt-1 compared with Ad-β-gal control (insert). (b) Dramatic reduction of sFlt-1 release from HUVEC 24 hours after transfection with sFlt-1 siRNA. (c) Cell proliferation after 48 hours in sFlt-1 siRNA transfected HUVEC. (d) Western blot showing VEGF receptor-2 (VEGFR-2) phosphorylation at tyrosine 951 in HUVEC transfected with sFlt-1, or control siRNA. VEGFR-2 and β-actin were used as a loading control. Data are expressed as mean (± SEM) or representative of three or more independent experiments performed in triplicate. ** P

Techniques Used: Infection, Western Blot, Transfection

23) Product Images from "The MARCH Family E3 Ubiquitin Ligase K5 Alters Monocyte Metabolism and Proliferation through Receptor Tyrosine Kinase Modulation"

Article Title: The MARCH Family E3 Ubiquitin Ligase K5 Alters Monocyte Metabolism and Proliferation through Receptor Tyrosine Kinase Modulation

Journal: PLoS Pathogens

doi: 10.1371/journal.ppat.1001331

THP-1 cells stably expressing K5 WT have sunitinib-sensitive increased RTK phosphorylation. ( A ) Normalized whole cell lysates from THP-1 cells, mock (PBS) or sunitinib (2 µM) treated, were subjected to western blot using an anti-phospho-tyrosine (4G10) (pTyr) antibody (top panel) and reprobed with an anti-actin antibody (bottom panel) to demonstrate equal loading. ( B ) Normalized WCL from parental or K5 WT-expressing THP-1 cells were subject to immunoprecipitation (IP) using anti-Flt-3 antibodies followed by western blot (WB) using an anti-phospho-tyrosine (4G10) (pTyr) antibody (top panel). The IP (middle panel)and whole cell lysates (WCL) (bottom panel) were probed with anti-Flt-3 antibodies, as controls. ( C ) 293T cells were co-transfected with expression constructs for K5 and Flt-4 or a Flt-4 mutant (G857R). Two days post-transfection, cell lysates were subjected to IP using anti-Flt-4 antibodies. western blot (WB) was performed using anti-phospho-tyrosine (4G10) (pTyr) antibodies (top panel) and re-probed with anti-Flt-4 antibodies (bottom panel). Data for all panels are representative of three independent experiments.
Figure Legend Snippet: THP-1 cells stably expressing K5 WT have sunitinib-sensitive increased RTK phosphorylation. ( A ) Normalized whole cell lysates from THP-1 cells, mock (PBS) or sunitinib (2 µM) treated, were subjected to western blot using an anti-phospho-tyrosine (4G10) (pTyr) antibody (top panel) and reprobed with an anti-actin antibody (bottom panel) to demonstrate equal loading. ( B ) Normalized WCL from parental or K5 WT-expressing THP-1 cells were subject to immunoprecipitation (IP) using anti-Flt-3 antibodies followed by western blot (WB) using an anti-phospho-tyrosine (4G10) (pTyr) antibody (top panel). The IP (middle panel)and whole cell lysates (WCL) (bottom panel) were probed with anti-Flt-3 antibodies, as controls. ( C ) 293T cells were co-transfected with expression constructs for K5 and Flt-4 or a Flt-4 mutant (G857R). Two days post-transfection, cell lysates were subjected to IP using anti-Flt-4 antibodies. western blot (WB) was performed using anti-phospho-tyrosine (4G10) (pTyr) antibodies (top panel) and re-probed with anti-Flt-4 antibodies (bottom panel). Data for all panels are representative of three independent experiments.

Techniques Used: Stable Transfection, Expressing, Western Blot, Immunoprecipitation, Transfection, Construct, Mutagenesis

K5 mediates rapid internalization of RTKs from the surface leading to increased signaling. ( A ) Equal numbers of the indicated THP-1 lines were mock or Dynasore (80 µM) treated for an hour at 37°C. Cells were then released at 37°C in serum free media for 0, 5, 10 or 30 minutes. At the indicated time points, cell aliquots were fixed and stained for surface levels of Flt-3, Flt-4 and PDGFR-ß followed by flow cytometric analysis. Representative experiments for each of the three receptors demonstrating the relative change in the expression of each RTK normalized to the fluorescence (geometric means) of mock treated samples is shown as line graphs, while the overall endocytosis rate of the receptor tyrosine kinases over the 30 minutes following dynasore release normalized to the time 0 point is shown as a bar graph with standard deviations. Line graph results are representative of at least three experiments ( B ) Equal numbers of vector- and K5 wt-expressing THP-1 were seeded in media containing 2% FBS. Following treatment with DMSO (mock) or Dynasore (80 µM) for 60 min and release in Dynasore-free medium at 37°C for 0, 30 or 60 minutes normalized whole cell lysates were prepared and subjected to western blot (WB) with an anti-phospho-tyrosine (4G10) (pTyr) antibody.
Figure Legend Snippet: K5 mediates rapid internalization of RTKs from the surface leading to increased signaling. ( A ) Equal numbers of the indicated THP-1 lines were mock or Dynasore (80 µM) treated for an hour at 37°C. Cells were then released at 37°C in serum free media for 0, 5, 10 or 30 minutes. At the indicated time points, cell aliquots were fixed and stained for surface levels of Flt-3, Flt-4 and PDGFR-ß followed by flow cytometric analysis. Representative experiments for each of the three receptors demonstrating the relative change in the expression of each RTK normalized to the fluorescence (geometric means) of mock treated samples is shown as line graphs, while the overall endocytosis rate of the receptor tyrosine kinases over the 30 minutes following dynasore release normalized to the time 0 point is shown as a bar graph with standard deviations. Line graph results are representative of at least three experiments ( B ) Equal numbers of vector- and K5 wt-expressing THP-1 were seeded in media containing 2% FBS. Following treatment with DMSO (mock) or Dynasore (80 µM) for 60 min and release in Dynasore-free medium at 37°C for 0, 30 or 60 minutes normalized whole cell lysates were prepared and subjected to western blot (WB) with an anti-phospho-tyrosine (4G10) (pTyr) antibody.

Techniques Used: Staining, Flow Cytometry, Expressing, Fluorescence, Plasmid Preparation, Western Blot

K5 interacts with and alters RTK localization. Equal cell numbers of the indicated THP-1 lines were fixed with paraformaldehyde (PFA) and ( A ) stained without permeabilization to determine surface expression or ( B ) permeabilized with saponin prior to staining to determine total expression of Flt-4, PDGFR-ß, Flt-3 and EGFR by flow cytometry. Data are representative of three independent experiments. ( B, Inset ) The relative ratio of surface versus total RTKs was determined for vector- and K5 WT-expressing THP-1 cells. ( C ) 293T cells were co-transfected with expression constructs for EGFR, PDGFR-ß, or Flt-4 and the indicated GST expression constructs. After two days, lysates were subjected to GST pull-down using glutathione-sepharose beads. Purified proteins and whole cell lysates (WCL) were subjected to western blot (WB) using anti-EGFR, -Flt-4 or -PDGFR-ß antibodies, followed by re-probing with anti-GST antibodies. Arrows indicate GST or GST-K5 WT and mutant specific bands. Data are representative of three independent experiments.
Figure Legend Snippet: K5 interacts with and alters RTK localization. Equal cell numbers of the indicated THP-1 lines were fixed with paraformaldehyde (PFA) and ( A ) stained without permeabilization to determine surface expression or ( B ) permeabilized with saponin prior to staining to determine total expression of Flt-4, PDGFR-ß, Flt-3 and EGFR by flow cytometry. Data are representative of three independent experiments. ( B, Inset ) The relative ratio of surface versus total RTKs was determined for vector- and K5 WT-expressing THP-1 cells. ( C ) 293T cells were co-transfected with expression constructs for EGFR, PDGFR-ß, or Flt-4 and the indicated GST expression constructs. After two days, lysates were subjected to GST pull-down using glutathione-sepharose beads. Purified proteins and whole cell lysates (WCL) were subjected to western blot (WB) using anti-EGFR, -Flt-4 or -PDGFR-ß antibodies, followed by re-probing with anti-GST antibodies. Arrows indicate GST or GST-K5 WT and mutant specific bands. Data are representative of three independent experiments.

Techniques Used: Staining, Expressing, Flow Cytometry, Cytometry, Plasmid Preparation, Transfection, Construct, Purification, Western Blot, Mutagenesis

24) Product Images from "Calcium dobesilate reduces VEGF signaling by interfering with heparan sulfate binding site and protects from vascular complications in diabetic mice"

Article Title: Calcium dobesilate reduces VEGF signaling by interfering with heparan sulfate binding site and protects from vascular complications in diabetic mice

Journal: PLoS ONE

doi: 10.1371/journal.pone.0218494

CaD inhibits VEGFR-2 phosphorylation and signaling in STZ diabetic mice. STZ mice were daily treated with vehicle, CaD or Enalapril and sacrificed at week 12 for the analysis. Non diabetic mice were used as control. Kidneys were isolated and homogenized as described in materials and methods. Phosphorylated VEGFR-2 (A) in the kidney lysates was determined by ELISA mean ± SD of 5–6 animals. Phosphorylated ERK1/2 (B) was analyzed by Western blot as described in Fig 1 . Vinculin was used as a loading control. Phosphorylated P38 (C) was analyzed by immunohistochemistry as described in the materials and methods section. Autofluorescence is shown in green, scale bar 100 μM. Representative image for n = 6/condition is depicted. *P
Figure Legend Snippet: CaD inhibits VEGFR-2 phosphorylation and signaling in STZ diabetic mice. STZ mice were daily treated with vehicle, CaD or Enalapril and sacrificed at week 12 for the analysis. Non diabetic mice were used as control. Kidneys were isolated and homogenized as described in materials and methods. Phosphorylated VEGFR-2 (A) in the kidney lysates was determined by ELISA mean ± SD of 5–6 animals. Phosphorylated ERK1/2 (B) was analyzed by Western blot as described in Fig 1 . Vinculin was used as a loading control. Phosphorylated P38 (C) was analyzed by immunohistochemistry as described in the materials and methods section. Autofluorescence is shown in green, scale bar 100 μM. Representative image for n = 6/condition is depicted. *P

Techniques Used: Mouse Assay, Isolation, Enzyme-linked Immunosorbent Assay, Western Blot, Immunohistochemistry

25) Product Images from "Up‐regulated basigin‐2 in microglia induced by hypoxia promotes retinal angiogenesis"

Article Title: Up‐regulated basigin‐2 in microglia induced by hypoxia promotes retinal angiogenesis

Journal: Journal of Cellular and Molecular Medicine

doi: 10.1111/jcmm.13256

Hypoxia up‐regulate basigin and HIF‐1α a expression in BV2 in vitro after 12, 24 hrs of treatment compared with untreated controls. Quantitative analysis of the expression of basigin and HIF‐1α by Western blotting under hypoxia after 12 or 24 hrs, with 0 hr as the baseline ( A ) and real‐time PCR analysis of the expression of basigin isoforms ( B ) in hypoxic BV2 cells ( n = 3). CT values in real‐time PCR analysis (basigin‐1 30.91 ± 1.80, basigin‐2 19.92 ± 0.76, β‐actin 12.56 ± 0.85) confirmed basigin‐1 expression was low. * P
Figure Legend Snippet: Hypoxia up‐regulate basigin and HIF‐1α a expression in BV2 in vitro after 12, 24 hrs of treatment compared with untreated controls. Quantitative analysis of the expression of basigin and HIF‐1α by Western blotting under hypoxia after 12 or 24 hrs, with 0 hr as the baseline ( A ) and real‐time PCR analysis of the expression of basigin isoforms ( B ) in hypoxic BV2 cells ( n = 3). CT values in real‐time PCR analysis (basigin‐1 30.91 ± 1.80, basigin‐2 19.92 ± 0.76, β‐actin 12.56 ± 0.85) confirmed basigin‐1 expression was low. * P

Techniques Used: Expressing, In Vitro, Western Blot, Real-time Polymerase Chain Reaction

Basigin was overexpressed in microglia, which accumulated in angiogenic sprouts in OIR. ( A ) Confocal images of IBA‐1(microglia marker) and CD31 stained retinal flat mounts in normoxic control mice (N) and OIR mice (OIR). ( B ) Confocal images of IBA‐1 and basigin stained retinal serial cryosections in two groups of mice (N and OIR). White arrowhead points to basigin + microglia associated with preretinal pathological tufts. GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer. The expression of basigin and HIF‐1α in the neural retina was analysed by Western blot. ** P
Figure Legend Snippet: Basigin was overexpressed in microglia, which accumulated in angiogenic sprouts in OIR. ( A ) Confocal images of IBA‐1(microglia marker) and CD31 stained retinal flat mounts in normoxic control mice (N) and OIR mice (OIR). ( B ) Confocal images of IBA‐1 and basigin stained retinal serial cryosections in two groups of mice (N and OIR). White arrowhead points to basigin + microglia associated with preretinal pathological tufts. GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer. The expression of basigin and HIF‐1α in the neural retina was analysed by Western blot. ** P

Techniques Used: Marker, Staining, Mouse Assay, Expressing, Western Blot

Related Articles

Flow Cytometry:

Article Title: Growth-Factor-Driven Rescue to Receptor Tyrosine Kinase (RTK) Inhibitors through Akt and Erk Phosphorylation in Pediatric Low Grade Astrocytoma and Ependymoma
Article Snippet: .. Flow cytometry analyses Cells were blocked by PBS 1% BSA (Bovine Serum Albumin, Sigma Aldrich, St Louis, MO, USA), and stained with anti-VEGFR-1 (Sigma Aldrich), anti-VEGFR-2-PE (Sigma Aldrich), anti-VEGFR-3-APC (R & D Systems, Minneapolis, MN, USA), anti-EGFR (Abcam, Cambridge, UK), anti-ErbB2-PE (R & D Systems), anti-ErbB3-APC (R & D Systems), anti-ErbB4 (R & D Systems), anti-HGFR/cMet-FITC (R & D Systems), anti-RON-APC (R & D Systems), anti-FGFR-1 (Cell Signaling, Danvers, MA, USA), anti-FGFR-2 (R & D Systems), anti-PDGFRα-biotin (BD Biosciences, San Jose, CA, USA), anti-PDGFRβ-PE (BD Biosciences). .. Primary FGFR-2, ErbB4 antibodies were visualized using a Rabbit anti-Mouse PE-conjugated secondary antibody (Dako Cytomation, Glostrup, Denmark), and FGFR-1 with a FITC-conjugated Swine anti-Rabbit secondary antibody (Dako Cytomation).

Cytometry:

Article Title: Growth-Factor-Driven Rescue to Receptor Tyrosine Kinase (RTK) Inhibitors through Akt and Erk Phosphorylation in Pediatric Low Grade Astrocytoma and Ependymoma
Article Snippet: .. Flow cytometry analyses Cells were blocked by PBS 1% BSA (Bovine Serum Albumin, Sigma Aldrich, St Louis, MO, USA), and stained with anti-VEGFR-1 (Sigma Aldrich), anti-VEGFR-2-PE (Sigma Aldrich), anti-VEGFR-3-APC (R & D Systems, Minneapolis, MN, USA), anti-EGFR (Abcam, Cambridge, UK), anti-ErbB2-PE (R & D Systems), anti-ErbB3-APC (R & D Systems), anti-ErbB4 (R & D Systems), anti-HGFR/cMet-FITC (R & D Systems), anti-RON-APC (R & D Systems), anti-FGFR-1 (Cell Signaling, Danvers, MA, USA), anti-FGFR-2 (R & D Systems), anti-PDGFRα-biotin (BD Biosciences, San Jose, CA, USA), anti-PDGFRβ-PE (BD Biosciences). .. Primary FGFR-2, ErbB4 antibodies were visualized using a Rabbit anti-Mouse PE-conjugated secondary antibody (Dako Cytomation, Glostrup, Denmark), and FGFR-1 with a FITC-conjugated Swine anti-Rabbit secondary antibody (Dako Cytomation).

Blocking Assay:

Article Title: VEGF-A/VEGFR-2 Signaling Leading to cAMP Response Element-Binding Protein Phosphorylation Is a Shared Pathway Underlying the Protective Effect of Preconditioning on Neurons and Endothelial Cells
Article Snippet: .. The sections were incubated with blocking solution containing primary antibodies at 4°C overnight and, on the next day, with secondary antibodies in blocking solution at room temperature for 1 h. The primary antibodies used were anti-phosphorylated CREB (pCREB) (1:100; Upstate), anti-VEGF-A (1:200; Millipore Bioscience Research Reagents), anti-VEGFR-1 (1:50; Millipore Bioscience Research Reagents), and anti-VEGFR-2 (1:100; Millipore Bioscience Research Reagents). ..

Article Title: VEGF-A/VEGFR-2 Signaling Leading to cAMP Response Element-Binding Protein Phosphorylation Is a Shared Pathway Underlying the Protective Effect of Preconditioning on Neurons and Endothelial Cells
Article Snippet: .. After blocking (1× PBS, 20% fetal bovine serum, 5% gelatin, and 0.1% Triton X-100) for 1 h, the slides were incubated overnight at 4°C with a mixture of two of the following primary antibodies: anti-CD31 (1:100; Bioscience Research Reagents), anti-VEGF-A (1:100; Millipore Bioscience Research Reagents or Santa Cruz Biotechnology), anti-VEGFR-1 (1:50; Millipore Bioscience Research Reagents), anti-VEGFR-2 (1:100; Millipore Bioscience Research Reagents), anti-pCREB (1:100; Upstate), anti-neuronal nuclear antigen (NeuN) (1:200; Millipore Bioscience Research Reagents), or anti-glial fibrillary acidic protein (GFAP) (1:400; Millipore Bioscience Research Reagents). .. The slides were washed three times with 0.1 m PBS and then incubated with Texas Red-conjugated anti-rabbit or anti-mouse IgG and FITC-conjugated anti-mouse, anti-rabbit, or anti-goat IgG (1:200 in blocking reagents; Jackson ImmunoResearch) for 1 h at room temperature.

Incubation:

Article Title: VEGF-A/VEGFR-2 Signaling Leading to cAMP Response Element-Binding Protein Phosphorylation Is a Shared Pathway Underlying the Protective Effect of Preconditioning on Neurons and Endothelial Cells
Article Snippet: .. The sections were incubated with blocking solution containing primary antibodies at 4°C overnight and, on the next day, with secondary antibodies in blocking solution at room temperature for 1 h. The primary antibodies used were anti-phosphorylated CREB (pCREB) (1:100; Upstate), anti-VEGF-A (1:200; Millipore Bioscience Research Reagents), anti-VEGFR-1 (1:50; Millipore Bioscience Research Reagents), and anti-VEGFR-2 (1:100; Millipore Bioscience Research Reagents). ..

Article Title: VEGF-A/VEGFR-2 Signaling Leading to cAMP Response Element-Binding Protein Phosphorylation Is a Shared Pathway Underlying the Protective Effect of Preconditioning on Neurons and Endothelial Cells
Article Snippet: .. After blocking (1× PBS, 20% fetal bovine serum, 5% gelatin, and 0.1% Triton X-100) for 1 h, the slides were incubated overnight at 4°C with a mixture of two of the following primary antibodies: anti-CD31 (1:100; Bioscience Research Reagents), anti-VEGF-A (1:100; Millipore Bioscience Research Reagents or Santa Cruz Biotechnology), anti-VEGFR-1 (1:50; Millipore Bioscience Research Reagents), anti-VEGFR-2 (1:100; Millipore Bioscience Research Reagents), anti-pCREB (1:100; Upstate), anti-neuronal nuclear antigen (NeuN) (1:200; Millipore Bioscience Research Reagents), or anti-glial fibrillary acidic protein (GFAP) (1:400; Millipore Bioscience Research Reagents). .. The slides were washed three times with 0.1 m PBS and then incubated with Texas Red-conjugated anti-rabbit or anti-mouse IgG and FITC-conjugated anti-mouse, anti-rabbit, or anti-goat IgG (1:200 in blocking reagents; Jackson ImmunoResearch) for 1 h at room temperature.

Western Blot:

Article Title: Intracrine Vascular Endothelial Growth Factor Signaling in Survival and Chemoresistance of Human Colorectal Cancer Cells
Article Snippet: .. Antibodies and suppliers for western blot analysis were as follows: anti-VEGF-A (R & D Systems, Minneapolis, MN, USA); anti-VEGFR-1 (Calbiochem, San Diego, CA, USA); anti-NRP-1 and anti-NRP-2 (Santa Cruz, CA, USA); anti-vinculin and anti-actin (Sigma-Aldrich); and anti-caspase-3, anti–cleaved caspase-3, anti-PARP, anti–cleaved PARP, anti-Bax and anti-survivin (Cell Signaling Technology, Danvers, MA, USA). .. Cell proliferation assay Cell growth was examined using an MTT assay (Sigma).

Staining:

Article Title: Growth-Factor-Driven Rescue to Receptor Tyrosine Kinase (RTK) Inhibitors through Akt and Erk Phosphorylation in Pediatric Low Grade Astrocytoma and Ependymoma
Article Snippet: .. Flow cytometry analyses Cells were blocked by PBS 1% BSA (Bovine Serum Albumin, Sigma Aldrich, St Louis, MO, USA), and stained with anti-VEGFR-1 (Sigma Aldrich), anti-VEGFR-2-PE (Sigma Aldrich), anti-VEGFR-3-APC (R & D Systems, Minneapolis, MN, USA), anti-EGFR (Abcam, Cambridge, UK), anti-ErbB2-PE (R & D Systems), anti-ErbB3-APC (R & D Systems), anti-ErbB4 (R & D Systems), anti-HGFR/cMet-FITC (R & D Systems), anti-RON-APC (R & D Systems), anti-FGFR-1 (Cell Signaling, Danvers, MA, USA), anti-FGFR-2 (R & D Systems), anti-PDGFRα-biotin (BD Biosciences, San Jose, CA, USA), anti-PDGFRβ-PE (BD Biosciences). .. Primary FGFR-2, ErbB4 antibodies were visualized using a Rabbit anti-Mouse PE-conjugated secondary antibody (Dako Cytomation, Glostrup, Denmark), and FGFR-1 with a FITC-conjugated Swine anti-Rabbit secondary antibody (Dako Cytomation).

Chloramphenicol Acetyltransferase Assay:

Article Title: A VEGF-A/SOX2/SRSF2 network controls VEGFR1 pre-mRNA alternative splicing in lung carcinoma cells
Article Snippet: .. The antibodies used were: anti-actin from Sigma, anti-tubulin (clone B512, sc-23948) from Santa Cruz, anti-SOX2 (AB5603) and anti-phospho-VEGFR1-Tyr1213 (cat07-758) from Millipore and anti-SRSF2 (clone 4F-11) from Euromedex. .. The specific anti-sVEGFR1-i13 was generated against a peptide mapping in the unique C-terminus .

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  • 96
    Millipore p vegfr2
    Correlation between (A) <t>pVEGFR2/VEGFR2</t> (p=0.007) and (B) EC pEGFR/EGFR (p=0.008) ratios and maximal reduction in tumor size (n=11). Complete responses were scored as −100% in tumor size change.
    P Vegfr2, supplied by Millipore, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Ligand-induced activation <t>VEGFR-1/-3</t> activation and downstream AKT and ERK activation by immunoblotting. A, SW480 cells cultured in serum-free media were pretreated with or without cediranib (100 nmol/L), were left untreated (SF; lanes 1, 6 ), or were
    P Vegfr 1, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore glp 1
    Effect of dietary fiber viscosity and fermentability on plasma <t>GLP-1</t> concentrations in the fasted state ( A ) and fed state ( B ). Values are mean ± SEM, n = 10–12 per group. Bars having different letters differ significantly ( p
    Glp 1, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 248 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Correlation between (A) pVEGFR2/VEGFR2 (p=0.007) and (B) EC pEGFR/EGFR (p=0.008) ratios and maximal reduction in tumor size (n=11). Complete responses were scored as −100% in tumor size change.

    Journal: The lancet oncology

    Article Title: A Phase II Study of Erlotinib and Bevacizumab in Patients with Recurrent or Metastatic Squamous Cell Carcinoma of the Head and Neck

    doi: 10.1016/S1470-2045(09)70002-6

    Figure Lengend Snippet: Correlation between (A) pVEGFR2/VEGFR2 (p=0.007) and (B) EC pEGFR/EGFR (p=0.008) ratios and maximal reduction in tumor size (n=11). Complete responses were scored as −100% in tumor size change.

    Article Snippet: Antibodies against human phosphorylated (p) VEGFR2 (PC460, Calbiochem;1:1,000), VEGFR2 (BD Pharmingen;1:100), pEGFR (Y1173, Santa Cruz Biotechnology;1:100), EGFR (1005, Santa Cruz;1:100), pERK (Calbiochem;1:100), ERK (Calbiochem1:100), pAKT (Calbiochem Cat# 1240011:100), AKT (Cell Signaling Cat# 2966;1:100) and CD31 (DakoCytomation1:100) were incubated with the rehydrated slides as previously described( ).

    Techniques:

    (A) Representative laser scanned images show high expression of phosphorylated VEGFR2/total VEGFR2 in CR compared to non-CR patients (pVEGFR2 stained red, VEGFR2 stained green). (B) High expression of phosphorylated EGFR/total EGFR in endothelial cells

    Journal: The lancet oncology

    Article Title: A Phase II Study of Erlotinib and Bevacizumab in Patients with Recurrent or Metastatic Squamous Cell Carcinoma of the Head and Neck

    doi: 10.1016/S1470-2045(09)70002-6

    Figure Lengend Snippet: (A) Representative laser scanned images show high expression of phosphorylated VEGFR2/total VEGFR2 in CR compared to non-CR patients (pVEGFR2 stained red, VEGFR2 stained green). (B) High expression of phosphorylated EGFR/total EGFR in endothelial cells

    Article Snippet: Antibodies against human phosphorylated (p) VEGFR2 (PC460, Calbiochem;1:1,000), VEGFR2 (BD Pharmingen;1:100), pEGFR (Y1173, Santa Cruz Biotechnology;1:100), EGFR (1005, Santa Cruz;1:100), pERK (Calbiochem;1:100), ERK (Calbiochem1:100), pAKT (Calbiochem Cat# 1240011:100), AKT (Cell Signaling Cat# 2966;1:100) and CD31 (DakoCytomation1:100) were incubated with the rehydrated slides as previously described( ).

    Techniques: Expressing, Staining

    Kaplan-Meier curves for the association between progression-free survival and (A) pVEGFR2/VEGFR2 (p=0.32) or (B) EC pEGFR/EGFR (p=0.12) ratios. Solid line represents subjects with a respective ratio at or above the median (n=6). Dotted line represents

    Journal: The lancet oncology

    Article Title: A Phase II Study of Erlotinib and Bevacizumab in Patients with Recurrent or Metastatic Squamous Cell Carcinoma of the Head and Neck

    doi: 10.1016/S1470-2045(09)70002-6

    Figure Lengend Snippet: Kaplan-Meier curves for the association between progression-free survival and (A) pVEGFR2/VEGFR2 (p=0.32) or (B) EC pEGFR/EGFR (p=0.12) ratios. Solid line represents subjects with a respective ratio at or above the median (n=6). Dotted line represents

    Article Snippet: Antibodies against human phosphorylated (p) VEGFR2 (PC460, Calbiochem;1:1,000), VEGFR2 (BD Pharmingen;1:100), pEGFR (Y1173, Santa Cruz Biotechnology;1:100), EGFR (1005, Santa Cruz;1:100), pERK (Calbiochem;1:100), ERK (Calbiochem1:100), pAKT (Calbiochem Cat# 1240011:100), AKT (Cell Signaling Cat# 2966;1:100) and CD31 (DakoCytomation1:100) were incubated with the rehydrated slides as previously described( ).

    Techniques:

    Ligand-induced activation VEGFR-1/-3 activation and downstream AKT and ERK activation by immunoblotting. A, SW480 cells cultured in serum-free media were pretreated with or without cediranib (100 nmol/L), were left untreated (SF; lanes 1, 6 ), or were

    Journal: Molecular cancer therapeutics

    Article Title: Targeting vascular endothelial growth factor receptor-1 and -3 with cediranib (AZD2171): effects on migration and invasion of gastrointestinal cancer cell lines

    doi: 10.1158/1535-7163.MCT-09-0380

    Figure Lengend Snippet: Ligand-induced activation VEGFR-1/-3 activation and downstream AKT and ERK activation by immunoblotting. A, SW480 cells cultured in serum-free media were pretreated with or without cediranib (100 nmol/L), were left untreated (SF; lanes 1, 6 ), or were

    Article Snippet: Phospho-specific antibodies were used to detect p-VEGFR-1 (Upstate, Millipore Corporation; 1:1,000), p-AKT, and p-ERK (Cell Signaling; 1:1,000) by standard immunoblotting methods as described above.

    Techniques: Activation Assay, Cell Culture

    Effect of dietary fiber viscosity and fermentability on plasma GLP-1 concentrations in the fasted state ( A ) and fed state ( B ). Values are mean ± SEM, n = 10–12 per group. Bars having different letters differ significantly ( p

    Journal: Nutrients

    Article Title: The Role of Viscosity and Fermentability of Dietary Fibers on Satiety- and Adiposity-Related Hormones in Rats

    doi: 10.3390/nu5062093

    Figure Lengend Snippet: Effect of dietary fiber viscosity and fermentability on plasma GLP-1 concentrations in the fasted state ( A ) and fed state ( B ). Values are mean ± SEM, n = 10–12 per group. Bars having different letters differ significantly ( p

    Article Snippet: There was a significant viscosity × fermentability interaction for plasma GLP-1 in the fasted state (p = 0.016).That is, the response of GLP-1 to fermentation was dependent on the viscosity. scFOS, the fermentable fiber with no viscosity, had the greatest plasma GLP-1 concentration, whereas in the diets containing fermentable fibers with some viscosity, GLP-1 concentrations were the lowest.

    Techniques: