Structured Review

Santa Cruz Biotechnology anti vegf c
sPLA 2 s induce the release of <t>VEGF-A</t> and <t>VEGF-C</t> from HLMs. A and B , HLMs were incubated (37°C, 24 h) with RPMI 1640 alone (Control) or with the indicated concentrations of hGIIA, hGX, or LPS. VEGF-A ( A ) and VEGF-C ( B ) release was determined by ELISA. Data are mean ± SEM of four experiments. * p
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1) Product Images from "Production of Vascular Endothelial Growth Factors from Human Lung Macrophages Induced by Group IIA and Group X Secreted Phospholipases A2"

Article Title: Production of Vascular Endothelial Growth Factors from Human Lung Macrophages Induced by Group IIA and Group X Secreted Phospholipases A2

Journal: Journal of immunology (Baltimore, Md. : 1950)

doi: 10.4049/jimmunol.0902501

sPLA 2 s induce the release of VEGF-A and VEGF-C from HLMs. A and B , HLMs were incubated (37°C, 24 h) with RPMI 1640 alone (Control) or with the indicated concentrations of hGIIA, hGX, or LPS. VEGF-A ( A ) and VEGF-C ( B ) release was determined by ELISA. Data are mean ± SEM of four experiments. * p
Figure Legend Snippet: sPLA 2 s induce the release of VEGF-A and VEGF-C from HLMs. A and B , HLMs were incubated (37°C, 24 h) with RPMI 1640 alone (Control) or with the indicated concentrations of hGIIA, hGX, or LPS. VEGF-A ( A ) and VEGF-C ( B ) release was determined by ELISA. Data are mean ± SEM of four experiments. * p

Techniques Used: Incubation, Enzyme-linked Immunosorbent Assay

HLMs constitutively express different forms of VEGF. A , Expression of VEGF mRNAs. RNA extraction from resting HLMs and RT-PCR was performed as described under Materials and Methods . Specific RT-PCR amplification products for VEGFA (isoforms 189, 165, and 121), VEGFB (isoforms 186 and 167), VEGFC, VEGFD, PlGF , and GAPDH were separated on 2% agarose gel, stained with ethidium bromide, and visualized with an image analysis system. The experiment shown is representative of three separate experiments. B , Detection of VEGF proteins. HLM protein extracts (40 μg per sample) were immunoblotted with anti–VEGF-A (gel I and II), anti-PlGF (gel III), anti–VEGF-B (gel IV), anti–VEGF-C (gel V), and anti–VEGF-D (gel VI) Abs. rhVEGF-A 165 , MCF-7 cells, EBNA expressing PlGF-1, RAW 264.7 cells were used as positive controls. Stripped membranes were reprobed with anti-GAPDH Ab to confirm equal protein content of each sample. Each Western blot shown is representative of three separate experiments.
Figure Legend Snippet: HLMs constitutively express different forms of VEGF. A , Expression of VEGF mRNAs. RNA extraction from resting HLMs and RT-PCR was performed as described under Materials and Methods . Specific RT-PCR amplification products for VEGFA (isoforms 189, 165, and 121), VEGFB (isoforms 186 and 167), VEGFC, VEGFD, PlGF , and GAPDH were separated on 2% agarose gel, stained with ethidium bromide, and visualized with an image analysis system. The experiment shown is representative of three separate experiments. B , Detection of VEGF proteins. HLM protein extracts (40 μg per sample) were immunoblotted with anti–VEGF-A (gel I and II), anti-PlGF (gel III), anti–VEGF-B (gel IV), anti–VEGF-C (gel V), and anti–VEGF-D (gel VI) Abs. rhVEGF-A 165 , MCF-7 cells, EBNA expressing PlGF-1, RAW 264.7 cells were used as positive controls. Stripped membranes were reprobed with anti-GAPDH Ab to confirm equal protein content of each sample. Each Western blot shown is representative of three separate experiments.

Techniques Used: Expressing, RNA Extraction, Reverse Transcription Polymerase Chain Reaction, Amplification, Agarose Gel Electrophoresis, Staining, Western Blot

2) Product Images from "Therapeutic Impact of Nanoparticle Therapy Targeting Tumor Associate Macrophages"

Article Title: Therapeutic Impact of Nanoparticle Therapy Targeting Tumor Associate Macrophages

Journal: Molecular cancer therapeutics

doi: 10.1158/1535-7163.MCT-17-0688

Depletion of TAMs is associated with a reduction in VEGF-C and downregulation of BRCA1 and BRCA2 gene expression
Figure Legend Snippet: Depletion of TAMs is associated with a reduction in VEGF-C and downregulation of BRCA1 and BRCA2 gene expression

Techniques Used: Expressing

3) Product Images from "Therapeutic Lymphangiogenesis With Implantation of Adipose-Derived Regenerative Cells"

Article Title: Therapeutic Lymphangiogenesis With Implantation of Adipose-Derived Regenerative Cells

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

doi: 10.1161/JAHA.112.000877

Implantation of ADRCs and lymphangiogenic cytokines in vivo. A, The abundance of VEGF‐C and HGF mRNA in the ADRC group was significantly greater than that of the control group by real‐time reverse transcriptase–polymerase chain reaction (VEGF‐C: 6.7‐fold, n=5, P
Figure Legend Snippet: Implantation of ADRCs and lymphangiogenic cytokines in vivo. A, The abundance of VEGF‐C and HGF mRNA in the ADRC group was significantly greater than that of the control group by real‐time reverse transcriptase–polymerase chain reaction (VEGF‐C: 6.7‐fold, n=5, P

Techniques Used: In Vivo, Polymerase Chain Reaction

The ability of ADRCs for lymphangiogenesis in vitro (functional assay and differentiation assay). A, Enzyme‐linked immunosorbent assay (ELISA) revealed that the concentration of VEGF‐C was upregulated in ADRC‐CM compared to control. B, rhVEGF‐C induced LEC migration in a dose‐dependent manner, and ADRC‐CM also induced LEC migration. HPF indicates high‐powered field. * P
Figure Legend Snippet: The ability of ADRCs for lymphangiogenesis in vitro (functional assay and differentiation assay). A, Enzyme‐linked immunosorbent assay (ELISA) revealed that the concentration of VEGF‐C was upregulated in ADRC‐CM compared to control. B, rhVEGF‐C induced LEC migration in a dose‐dependent manner, and ADRC‐CM also induced LEC migration. HPF indicates high‐powered field. * P

Techniques Used: In Vitro, Functional Assay, Differentiation Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay, Migration

Newly established mouse model of tail lymphedema. A, Circumferential annulus of skin (2 mm wide) located 10 mm distal to the tail base, excluding a 4‐mm 2 dermal flap at the ventral side, was removed from the tail. Good blood perfusion was maintained at distal site. B, Tail lymphedema was induced within a few days after the procedure, and the tail diameter peaked at around postoperative day 7. Lymphedema continued in the PBS group to at least 28 days. C, Representative photomicrographs of histological sections in lymphedematous tail tissues distal to the incision site. The space of subcutaneous tissues was edematous and dilated in the PBS group, but the space was almost normal in the ADRC implantation group (double‐headed arrow). Scale bar=2 mm. D, As a positive control, a marked reduction of the edema was observed after day 11 in VEGF‐C protein–administered group compared to PBS group ( P
Figure Legend Snippet: Newly established mouse model of tail lymphedema. A, Circumferential annulus of skin (2 mm wide) located 10 mm distal to the tail base, excluding a 4‐mm 2 dermal flap at the ventral side, was removed from the tail. Good blood perfusion was maintained at distal site. B, Tail lymphedema was induced within a few days after the procedure, and the tail diameter peaked at around postoperative day 7. Lymphedema continued in the PBS group to at least 28 days. C, Representative photomicrographs of histological sections in lymphedematous tail tissues distal to the incision site. The space of subcutaneous tissues was edematous and dilated in the PBS group, but the space was almost normal in the ADRC implantation group (double‐headed arrow). Scale bar=2 mm. D, As a positive control, a marked reduction of the edema was observed after day 11 in VEGF‐C protein–administered group compared to PBS group ( P

Techniques Used: Positive Control

Possible mechanisms of lymphangiogenesis mediated by ADRC implantation. We propose 2 main mechanisms: First, implanted ADRCs release cytokines, including VEGF‐C, that might stimulate migration and proliferation of residual LECs and eventual lymphangiogenesis. Second, cytokines released from ADRCs could augment mobilization and/or recruitment of bone marrow–derived M2 macrophages serving as lymphatic endothelial progenitors. There is little evidence that implanted ADRCs directly transdifferentiate into mature LECs.
Figure Legend Snippet: Possible mechanisms of lymphangiogenesis mediated by ADRC implantation. We propose 2 main mechanisms: First, implanted ADRCs release cytokines, including VEGF‐C, that might stimulate migration and proliferation of residual LECs and eventual lymphangiogenesis. Second, cytokines released from ADRCs could augment mobilization and/or recruitment of bone marrow–derived M2 macrophages serving as lymphatic endothelial progenitors. There is little evidence that implanted ADRCs directly transdifferentiate into mature LECs.

Techniques Used: Migration, Derivative Assay

4) Product Images from "Grafting and Early Expression of Growth Factors from Adipose-Derived Stem Cells Transplanted into the Cochlea, in a Guinea Pig Model of Acoustic Trauma"

Article Title: Grafting and Early Expression of Growth Factors from Adipose-Derived Stem Cells Transplanted into the Cochlea, in a Guinea Pig Model of Acoustic Trauma

Journal: Frontiers in Cellular Neuroscience

doi: 10.3389/fncel.2014.00334

VEGFs expression in the cochlea . Representative images from confocal microscopy of cochlear cryosections collected at day 7 after surgery: (A) VEGF-A staining (red fluorescence) in control unexposed cochlea; (B) green fluorescent cells in the scala media (arrow head); (C) VEGF-A expression in noise-exposed; (D) increased VEGF-A expression in the scala media and in implanted ASCs near the stria vascularis (arrow head) and in the spiral ganglion (arrow); (E–H) higher magnification of the stria vascularis ; and (I–K) of the spiral ganglion; (L–N) VEGF-C and GFP fluorescence in cochlear sections; ASCs close to the lateral wall (arrows). Panel (O) shows electrophoretic bands of cDNA migration, from rtPCR analysis performed in cultured ASCs. VEGF-A, vascular endothelial growth factor A; VEGF-C, vascular endothelial growth factor C.
Figure Legend Snippet: VEGFs expression in the cochlea . Representative images from confocal microscopy of cochlear cryosections collected at day 7 after surgery: (A) VEGF-A staining (red fluorescence) in control unexposed cochlea; (B) green fluorescent cells in the scala media (arrow head); (C) VEGF-A expression in noise-exposed; (D) increased VEGF-A expression in the scala media and in implanted ASCs near the stria vascularis (arrow head) and in the spiral ganglion (arrow); (E–H) higher magnification of the stria vascularis ; and (I–K) of the spiral ganglion; (L–N) VEGF-C and GFP fluorescence in cochlear sections; ASCs close to the lateral wall (arrows). Panel (O) shows electrophoretic bands of cDNA migration, from rtPCR analysis performed in cultured ASCs. VEGF-A, vascular endothelial growth factor A; VEGF-C, vascular endothelial growth factor C.

Techniques Used: Expressing, Confocal Microscopy, Staining, Fluorescence, Migration, Reverse Transcription Polymerase Chain Reaction, Cell Culture

5) Product Images from "MT1-MMP sheds LYVE-1 on lymphatic endothelial cells and suppresses VEGF-C production to inhibit lymphangiogenesis"

Article Title: MT1-MMP sheds LYVE-1 on lymphatic endothelial cells and suppresses VEGF-C production to inhibit lymphangiogenesis

Journal: Nature Communications

doi: 10.1038/ncomms10824

Blocking VEGFR3 activities inhibits corneal lymphangiogenesis in Mmp14 −/− mice. Real-time qPCR analyses of mRNA for Vegf-c ( a ), Vegf-a ( b ), Vegf-d ( c ), Tnf-α ( d ), Il-1β ( e ), Mcp-1 ( f ) and Mip-2 ( g ) in WT and Mmp14 −/− corneas at different ages (P3, P8 and P15) (* P
Figure Legend Snippet: Blocking VEGFR3 activities inhibits corneal lymphangiogenesis in Mmp14 −/− mice. Real-time qPCR analyses of mRNA for Vegf-c ( a ), Vegf-a ( b ), Vegf-d ( c ), Tnf-α ( d ), Il-1β ( e ), Mcp-1 ( f ) and Mip-2 ( g ) in WT and Mmp14 −/− corneas at different ages (P3, P8 and P15) (* P

Techniques Used: Blocking Assay, Mouse Assay, Real-time Polymerase Chain Reaction

MT1-MMP restrains VEGF-C production in macrophages. ( a ) Real-time qPCR analyses of Vegf-c transcripts in BMMs from WT and Mmp14 −/− mice on LPS (1 μg ml −1 ) stimulation (** P
Figure Legend Snippet: MT1-MMP restrains VEGF-C production in macrophages. ( a ) Real-time qPCR analyses of Vegf-c transcripts in BMMs from WT and Mmp14 −/− mice on LPS (1 μg ml −1 ) stimulation (** P

Techniques Used: Real-time Polymerase Chain Reaction, Mouse Assay

MT1-MMP suppresses VEGF-C expression via regulation of PI3Kδ signalling. ( a ) Western blot analyses of VEGF-C in WT and Mmp14 −/− BMMs with ectopic expression of control vector, full-length MT1-MMP, the cytosolic domain-deleted MT1-MMP or the E/A240 mutant MT1-MMP after stimulation with 1 μg ml −1 LPS for 24 h. ( b ) qPCR analyses of Vegf-c mRNA expression in WT and Mmp14 −/− BMMs with ectopic expression of a control vector, full-length MT1-MMP, the cytosolic domain-deleted or the E/A240 mutant MT1-MMP. Cells were stimulated with 1 μg ml −1 LPS for 6 h before the analyses (** P
Figure Legend Snippet: MT1-MMP suppresses VEGF-C expression via regulation of PI3Kδ signalling. ( a ) Western blot analyses of VEGF-C in WT and Mmp14 −/− BMMs with ectopic expression of control vector, full-length MT1-MMP, the cytosolic domain-deleted MT1-MMP or the E/A240 mutant MT1-MMP after stimulation with 1 μg ml −1 LPS for 24 h. ( b ) qPCR analyses of Vegf-c mRNA expression in WT and Mmp14 −/− BMMs with ectopic expression of a control vector, full-length MT1-MMP, the cytosolic domain-deleted or the E/A240 mutant MT1-MMP. Cells were stimulated with 1 μg ml −1 LPS for 6 h before the analyses (** P

Techniques Used: Expressing, Western Blot, Plasmid Preparation, Mutagenesis, Real-time Polymerase Chain Reaction

6) Product Images from "VEGFR3 inhibition chemosensitizes lung adenocarcinoma A549 cells in the tumor-associated macrophage microenvironment through upregulation of p53 and PTEN"

Article Title: VEGFR3 inhibition chemosensitizes lung adenocarcinoma A549 cells in the tumor-associated macrophage microenvironment through upregulation of p53 and PTEN

Journal: Oncology Reports

doi: 10.3892/or.2017.5969

Expression levels of VEGF-C, VEGF-D, and VEGFR3 in the lung adenocarcinoma A549 cell line. (A) RT-PCR evaluations of VEGF-C mRNA, VEGF-D mRNA and VEGFR3 mRNA in co-cultured M2 macrophages (CO-M2) and co-cultured A549 cells (CO-A549) are presented. (B) IHC demonstrating that VEGFR3 expression in human lung adenocarcinoma is primarily found in tumor cells than in stromal and adjacent cells. VEGFR3 expression was also found in blood vessels. Scale bars, 20 µm. (C) Semi-quantitative RT-PCR and qRT-PCR evaluation of VEGF-C mRNA, VEGF-D mRNA and VEGFR3 mRNA are presented. The co-cultured A549 cells (CO-A549) had a higher expression level of VEGF-C mRNA and VEGF-D mRNA than that of the A549 cells alone. (D) Western blot analysis confirmed a high protein expression of VEGF-C and VEGFR3 in CO-A549 cells. *P
Figure Legend Snippet: Expression levels of VEGF-C, VEGF-D, and VEGFR3 in the lung adenocarcinoma A549 cell line. (A) RT-PCR evaluations of VEGF-C mRNA, VEGF-D mRNA and VEGFR3 mRNA in co-cultured M2 macrophages (CO-M2) and co-cultured A549 cells (CO-A549) are presented. (B) IHC demonstrating that VEGFR3 expression in human lung adenocarcinoma is primarily found in tumor cells than in stromal and adjacent cells. VEGFR3 expression was also found in blood vessels. Scale bars, 20 µm. (C) Semi-quantitative RT-PCR and qRT-PCR evaluation of VEGF-C mRNA, VEGF-D mRNA and VEGFR3 mRNA are presented. The co-cultured A549 cells (CO-A549) had a higher expression level of VEGF-C mRNA and VEGF-D mRNA than that of the A549 cells alone. (D) Western blot analysis confirmed a high protein expression of VEGF-C and VEGFR3 in CO-A549 cells. *P

Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Cell Culture, Immunohistochemistry, Quantitative RT-PCR, Western Blot

VEGFR3 inhibition improves p53 and PTEN protein expression through p-ERK. (A) Western blot analysis of phosphoproteins, control, MAZ51 and VEGF-C-treated co-cultured A549 cells revealed that MAZ51 treatment was associated with decreased p-ERK. (B) Western blot anlaysis demonstrating that MAZ51 treatment is associated with upregulated proteins p53 and PTEN in co-cultured A549 cells. (C) MEK1 inhibition (U0126) is associated with upregulation of p53 and PTEN mRNA levels. **P
Figure Legend Snippet: VEGFR3 inhibition improves p53 and PTEN protein expression through p-ERK. (A) Western blot analysis of phosphoproteins, control, MAZ51 and VEGF-C-treated co-cultured A549 cells revealed that MAZ51 treatment was associated with decreased p-ERK. (B) Western blot anlaysis demonstrating that MAZ51 treatment is associated with upregulated proteins p53 and PTEN in co-cultured A549 cells. (C) MEK1 inhibition (U0126) is associated with upregulation of p53 and PTEN mRNA levels. **P

Techniques Used: Inhibition, Expressing, Western Blot, Cell Culture

7) Product Images from "VEGF-A, VEGF-C, and VEGF-D in Colorectal Cancer Progression"

Article Title: VEGF-A, VEGF-C, and VEGF-D in Colorectal Cancer Progression

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

doi:

mRNA expression of VEGF-A, VEGF-C, and VEGF-D through the adenoma-carcinoma sequence. Median values with interquartile range (box) and range (whiskers).
Figure Legend Snippet: mRNA expression of VEGF-A, VEGF-C, and VEGF-D through the adenoma-carcinoma sequence. Median values with interquartile range (box) and range (whiskers).

Techniques Used: Expressing, Sequencing

8) Product Images from "Significance of Tumor-Associated Stroma in Promotion of Intratumoral Lymphangiogenesis "

Article Title: Significance of Tumor-Associated Stroma in Promotion of Intratumoral Lymphangiogenesis

Journal: The American Journal of Pathology

doi: 10.2353/ajpath.2008.070360

Increased Expression of VEGF-C and -D in Has2-Overexpressing Tumors
Figure Legend Snippet: Increased Expression of VEGF-C and -D in Has2-Overexpressing Tumors

Techniques Used: Expressing

Expression and tissue localization of lymphangiogenic factors. A: Relative mRNA expression of VEGF-C and -D in mammary tumor cells. Total RNA from Has2 +Neo and Has2 ΔNeo tumors were transcribed into cDNA and the relative mRNA levels of
Figure Legend Snippet: Expression and tissue localization of lymphangiogenic factors. A: Relative mRNA expression of VEGF-C and -D in mammary tumor cells. Total RNA from Has2 +Neo and Has2 ΔNeo tumors were transcribed into cDNA and the relative mRNA levels of

Techniques Used: Expressing

9) Product Images from "Lack of Lymphatics and Lymph Node-Mediated Immunity in Choroidal Neovascularization"

Article Title: Lack of Lymphatics and Lymph Node-Mediated Immunity in Choroidal Neovascularization

Journal: Investigative Ophthalmology & Visual Science

doi: 10.1167/iovs.12-10341

VEGF-C and VEGFR-3 expression in mouse CNV model. ( A ) Representative Western blot samples from lasered and normal control choroids–RPE complex (days 3 and 7) of mice with α-VEGFR-2, VEGFR-3, and VEGF-C antibodies, showing upregulation
Figure Legend Snippet: VEGF-C and VEGFR-3 expression in mouse CNV model. ( A ) Representative Western blot samples from lasered and normal control choroids–RPE complex (days 3 and 7) of mice with α-VEGFR-2, VEGFR-3, and VEGF-C antibodies, showing upregulation

Techniques Used: Expressing, Western Blot, Mouse Assay

10) Product Images from "The secretory proprotein convertases furin, PC5, and PC7 activate VEGF-C to induce tumorigenesis"

Article Title: The secretory proprotein convertases furin, PC5, and PC7 activate VEGF-C to induce tumorigenesis

Journal: Journal of Clinical Investigation

doi: 10.1172/JCI200317220

Endogenous VEGF-C processing by PC-like activity and coexpression of furin and VEGF-C in mouse tissues. ( a ) Endogenous proVEGF-C processing was analyzed by Western blotting of PC3 cell–conditioned media obtained from cells transiently transfected with either the pIRES2-EGFP empty vectors (Control) or vector expressing α1-antitrypsin, pSKI-1, α1-PDX, p-furin, furin, or PC5. The corresponding percentages of proVEGF-C cleavage calculated from the ratio of band intensities of VEGF-C/(proVEGF-C + VEGF-C) are indicated. ( b ) Total RNA was extracted from PC3 cells and the indicated tissues and organs, and RT-PCR analysis was performed using primers specific for VEGF-C, furin, and GAPDH (control) under the conditions described in Methods. Results shown are representative of three experiments.
Figure Legend Snippet: Endogenous VEGF-C processing by PC-like activity and coexpression of furin and VEGF-C in mouse tissues. ( a ) Endogenous proVEGF-C processing was analyzed by Western blotting of PC3 cell–conditioned media obtained from cells transiently transfected with either the pIRES2-EGFP empty vectors (Control) or vector expressing α1-antitrypsin, pSKI-1, α1-PDX, p-furin, furin, or PC5. The corresponding percentages of proVEGF-C cleavage calculated from the ratio of band intensities of VEGF-C/(proVEGF-C + VEGF-C) are indicated. ( b ) Total RNA was extracted from PC3 cells and the indicated tissues and organs, and RT-PCR analysis was performed using primers specific for VEGF-C, furin, and GAPDH (control) under the conditions described in Methods. Results shown are representative of three experiments.

Techniques Used: Activity Assay, Western Blot, Transfection, Plasmid Preparation, Expressing, Reverse Transcription Polymerase Chain Reaction

In vitro digestions of Q-h-VEGF-C with recombinant furin, PC5, and PC7. ( a ) RP-HPLC chromatogram of the crude digest following 4 hours of incubation at 37°C of 20 μg of QVEGF-C with furin, PC5, or PC7 in 25 mM Tris, 25 mM Mes, and 2.5 mM CaCl 2 , pH 7.4. The elution of the peaks was monitored on-line by UV absorbance at 214 nm as well as by fluorescence detectors (λ ex , 320 nm; λ em , 420 nm). ( b ) MALDI-ToF mass spectra of the crude digests following 24 hours of incubation at 37°C of 20 μg of QVEGF-C with furin, PC5, and PC7 in 25 mM Tris, 25 mM Mes, and 2.5 mM CaCl 2 , pH 7.4. Note the absence of the peak at m/z 1,703 suggesting complete cleavage of QVEGF-C. The peaks at m/z 1,129 and 595 were attributed to the highly fluorescent N-terminal (NT) (Abz-Q-VHSIIRR-OH) and the nonfluorescent C-terminal (CT) [SLP(NO 2 )-A-CONH 2 ] fragments, respectively. The peaks at m/z 972 and 663 are not PC-dependent since they are also present in the crude digest of QVEGF-C by wild-type medium (data not shown). NT-R, the N-terminal sequence without Arginine residue.
Figure Legend Snippet: In vitro digestions of Q-h-VEGF-C with recombinant furin, PC5, and PC7. ( a ) RP-HPLC chromatogram of the crude digest following 4 hours of incubation at 37°C of 20 μg of QVEGF-C with furin, PC5, or PC7 in 25 mM Tris, 25 mM Mes, and 2.5 mM CaCl 2 , pH 7.4. The elution of the peaks was monitored on-line by UV absorbance at 214 nm as well as by fluorescence detectors (λ ex , 320 nm; λ em , 420 nm). ( b ) MALDI-ToF mass spectra of the crude digests following 24 hours of incubation at 37°C of 20 μg of QVEGF-C with furin, PC5, and PC7 in 25 mM Tris, 25 mM Mes, and 2.5 mM CaCl 2 , pH 7.4. Note the absence of the peak at m/z 1,703 suggesting complete cleavage of QVEGF-C. The peaks at m/z 1,129 and 595 were attributed to the highly fluorescent N-terminal (NT) (Abz-Q-VHSIIRR-OH) and the nonfluorescent C-terminal (CT) [SLP(NO 2 )-A-CONH 2 ] fragments, respectively. The peaks at m/z 972 and 663 are not PC-dependent since they are also present in the crude digest of QVEGF-C by wild-type medium (data not shown). NT-R, the N-terminal sequence without Arginine residue.

Techniques Used: In Vitro, Recombinant, High Performance Liquid Chromatography, Incubation, Fluorescence, Sequencing

Blockade of proVEGF-C processing inhibits in vivo tumor cell growth. ( a ) Populations of control CHO cells (Ctl) or CHO cells expressing wild-type (VEGF-C) or mutant VEGF-C (VEGF-C/mut) were injected subcutaneously into 4-week-old male nude mice. Tumor size was measured every 3 days. ( b ) Starved populations of control CHO cells or CHO cells expressing wild-type (VEGF-C) or mutant VEGF-C (VEGF-C/mut) were incubated for 24 hours in medium containing increasing concentrations of serum (0–10% FCS). [ 3 H]thymidine was added for the final 6 hours of incubation. [ 3 ). Data are presented as mean ± SE of four experiments.
Figure Legend Snippet: Blockade of proVEGF-C processing inhibits in vivo tumor cell growth. ( a ) Populations of control CHO cells (Ctl) or CHO cells expressing wild-type (VEGF-C) or mutant VEGF-C (VEGF-C/mut) were injected subcutaneously into 4-week-old male nude mice. Tumor size was measured every 3 days. ( b ) Starved populations of control CHO cells or CHO cells expressing wild-type (VEGF-C) or mutant VEGF-C (VEGF-C/mut) were incubated for 24 hours in medium containing increasing concentrations of serum (0–10% FCS). [ 3 H]thymidine was added for the final 6 hours of incubation. [ 3 ). Data are presented as mean ± SE of four experiments.

Techniques Used: In Vivo, CTL Assay, Expressing, Mutagenesis, Injection, Mouse Assay, Incubation

Processing of proVEGF-C by furin, PC5, and PC7. ( a ) Schematic representation of the primary structure of the 419-AA human proVEGF-C. Shown are the signal peptide (SP), PC-processing site (HSIIRR 227 SL), an unknown protease site (indicated by question mark) that generates the 21-kDa VEGF-C, and the Flag attached to the C-terminus. ProVEGF-C processing was analyzed by biosynthesis ( b ) and Western blotting ( c ) of LoVo-C5–conditioned media obtained from cells transiently transfected with either the empty vectors (None), pIRES2-EGFP vector and pcDNA3-zeo-Flag.cm5 vector containing proVEGF-C (Control), or with the pcDNA3-zeo-Flag.cm5 vector containing proVEGF-C and pIRES2-EGFP vector that expresses full-length human furin, PACE4, or SKI-1; mouse PC5A or PC5B; or rat PC7. The corresponding percentages of proVEGF-C cleavage calculated from the ratio of band intensities of VEGF-C/(proVEGF-C + VEGF-C) are indicated.
Figure Legend Snippet: Processing of proVEGF-C by furin, PC5, and PC7. ( a ) Schematic representation of the primary structure of the 419-AA human proVEGF-C. Shown are the signal peptide (SP), PC-processing site (HSIIRR 227 SL), an unknown protease site (indicated by question mark) that generates the 21-kDa VEGF-C, and the Flag attached to the C-terminus. ProVEGF-C processing was analyzed by biosynthesis ( b ) and Western blotting ( c ) of LoVo-C5–conditioned media obtained from cells transiently transfected with either the empty vectors (None), pIRES2-EGFP vector and pcDNA3-zeo-Flag.cm5 vector containing proVEGF-C (Control), or with the pcDNA3-zeo-Flag.cm5 vector containing proVEGF-C and pIRES2-EGFP vector that expresses full-length human furin, PACE4, or SKI-1; mouse PC5A or PC5B; or rat PC7. The corresponding percentages of proVEGF-C cleavage calculated from the ratio of band intensities of VEGF-C/(proVEGF-C + VEGF-C) are indicated.

Techniques Used: Western Blot, Transfection, Plasmid Preparation

11) Product Images from "MULTIMERIN2 binds VEGF-A primarily via the carbohydrate chains exerting an angiostatic function and impairing tumor growth"

Article Title: MULTIMERIN2 binds VEGF-A primarily via the carbohydrate chains exerting an angiostatic function and impairing tumor growth

Journal: Oncotarget

doi:

MMRN2 specifically binds to VEGF-A A. Dose response plot of the interaction of MMRN2 with VEGF-A 165 and VEGF-A 121 , as obtained by surface plasmon resonance. B. Dose response plot of the interaction of MMRN2 with VEGF-A 165, VEGF-A 145 and VEGF-A 189 , as obtained by surface plasmon resonance. C, D, E. Sensograms reporting the binding of VEGF-C, VEGF-D and PlGF-1, as assessed by surface plasmon resonance. F. Sensogram of the comparison of the binding of VEGF-A 165 , VEGF-C, VEGF-D and PlGF-1 at the concentration of 200nM to MMRN2 as assessed by surface plasmon resonance. G. Dose response plot of the interaction of MMRN2 with VEGF-A 165 , VEGF-C, VEGF-D and PlGF-1 as obtained by surface plasmon resonance. All experiments were repeated at least three times.
Figure Legend Snippet: MMRN2 specifically binds to VEGF-A A. Dose response plot of the interaction of MMRN2 with VEGF-A 165 and VEGF-A 121 , as obtained by surface plasmon resonance. B. Dose response plot of the interaction of MMRN2 with VEGF-A 165, VEGF-A 145 and VEGF-A 189 , as obtained by surface plasmon resonance. C, D, E. Sensograms reporting the binding of VEGF-C, VEGF-D and PlGF-1, as assessed by surface plasmon resonance. F. Sensogram of the comparison of the binding of VEGF-A 165 , VEGF-C, VEGF-D and PlGF-1 at the concentration of 200nM to MMRN2 as assessed by surface plasmon resonance. G. Dose response plot of the interaction of MMRN2 with VEGF-A 165 , VEGF-C, VEGF-D and PlGF-1 as obtained by surface plasmon resonance. All experiments were repeated at least three times.

Techniques Used: SPR Assay, Binding Assay, Concentration Assay

12) Product Images from "Hypoxia inducible factor-1? correlates with vascular endothelial growth factor A and C indicating worse prognosis in clear cell renal cell carcinoma"

Article Title: Hypoxia inducible factor-1? correlates with vascular endothelial growth factor A and C indicating worse prognosis in clear cell renal cell carcinoma

Journal: Journal of Experimental & Clinical Cancer Research : CR

doi: 10.1186/1756-9966-28-40

Kaplan-Meier cumulative survival analysis according to staining for nuclear and cytoplasmic HIF-1α, VEGF-A and VEGF-C . The log-rank test showed significantly shorter overall survival in patients with tumors showing low nHIF-1α (p = 0.005) (A) and low pVEGF-C (p = 0.008) (D). The 5-year survival rate was 32% for patients whose tumors showed low nHIF-1α vs . 65% for patients whose tumors showed high nHIF-1α (A); and 40% for patients whose tumors showed low pVEGF-C vs . 61% for patients whose tumors showed high pVEGF-C (D). The log-rank test showed significantly shorter overall survival in patients with tumors showing high cHIF-1α (p = 0.018) (B) and high dVEGF-A (p = 0.024) (C). The 5-year survival rate was 60% for patients whose tumors showed low cHIF-1α vs . 40% for patients whose tumors showed high cHIF-1α (B); and 59% for patients whose tumors showed low dVEGF-A vs . 40% for patients whose tumors showed high dVEGF-A (C).
Figure Legend Snippet: Kaplan-Meier cumulative survival analysis according to staining for nuclear and cytoplasmic HIF-1α, VEGF-A and VEGF-C . The log-rank test showed significantly shorter overall survival in patients with tumors showing low nHIF-1α (p = 0.005) (A) and low pVEGF-C (p = 0.008) (D). The 5-year survival rate was 32% for patients whose tumors showed low nHIF-1α vs . 65% for patients whose tumors showed high nHIF-1α (A); and 40% for patients whose tumors showed low pVEGF-C vs . 61% for patients whose tumors showed high pVEGF-C (D). The log-rank test showed significantly shorter overall survival in patients with tumors showing high cHIF-1α (p = 0.018) (B) and high dVEGF-A (p = 0.024) (C). The 5-year survival rate was 60% for patients whose tumors showed low cHIF-1α vs . 40% for patients whose tumors showed high cHIF-1α (B); and 59% for patients whose tumors showed low dVEGF-A vs . 40% for patients whose tumors showed high dVEGF-A (C).

Techniques Used: Staining

Immunohistochemical staining of HIF-1α, VEGF-A and VEGF-C in normal renal tissue (A-C) and clear cell renal cell carcinoma (CCRCC) (D-F) . A homogeneous cytoplasmic staining of tubular cells and weak staining in glomerules was observed with HIF-1α (A), while VEGF-A and VEGF-C were positive in tubular cells, glomerular mesangium and interstitial macrophages (B and C). In CCRCC, HIF-1α immmunoreactivity was nuclear and/or cytoplasmic (D), while it was perimembranous and/or diffuse cytoplasmic for VEGF-A and VEFG-C (E and F). (magnification ×200).
Figure Legend Snippet: Immunohistochemical staining of HIF-1α, VEGF-A and VEGF-C in normal renal tissue (A-C) and clear cell renal cell carcinoma (CCRCC) (D-F) . A homogeneous cytoplasmic staining of tubular cells and weak staining in glomerules was observed with HIF-1α (A), while VEGF-A and VEGF-C were positive in tubular cells, glomerular mesangium and interstitial macrophages (B and C). In CCRCC, HIF-1α immmunoreactivity was nuclear and/or cytoplasmic (D), while it was perimembranous and/or diffuse cytoplasmic for VEGF-A and VEFG-C (E and F). (magnification ×200).

Techniques Used: Immunohistochemistry, Staining

13) Product Images from "CYR61 (CCN1) Is Essential for Placental Development and Vascular Integrity"

Article Title: CYR61 (CCN1) Is Essential for Placental Development and Vascular Integrity

Journal: Molecular and Cellular Biology

doi: 10.1128/MCB.22.24.8709-8720.2002

(A) β-Galactosidase staining was detected at the chorioallantoic junction at E10.5, when embryonic vessels were actively bifurcating and invaded the placenta. Intense staining was found in the allantoic mesoderm and inner endothelial lining associated with bifurcating vessels (arrowhead). (B) By E13.5, β-galactosidase staining was localized to the vessels in the chorionic plate, maternal decidua, and trophoblast giant cells. (C) Higher magnification of panel B is given to demonstrate staining in giant cells (arrowheads). (D) VEGF-C was detected in the allantoic mesoderm (arrowheads) and chorion of WT E9.5 placenta by immunostaining but not in allantoic mesoderm of Cyr61 −/− placenta (E). (F) In Northern blot analysis, serum-starved mouse embryonic fibroblasts isolated from E14.5 WT embryos were treated with purified recombinant CYR61 for various durations before the total RNA was isolated. Vegf-C mRNA was upregulated sixfold after 6 h of CYR61 treatment, and this upregulation was sustained for at least 24 h. The hybridization signal was quantified by Phosphorimager. Al, allantois; Ch, chorion; De, decidua; G, giant cells; La, labyrinth; Sp, spongiotrophoblast; U, umbilical cord. Yellow bars, 50 μm.
Figure Legend Snippet: (A) β-Galactosidase staining was detected at the chorioallantoic junction at E10.5, when embryonic vessels were actively bifurcating and invaded the placenta. Intense staining was found in the allantoic mesoderm and inner endothelial lining associated with bifurcating vessels (arrowhead). (B) By E13.5, β-galactosidase staining was localized to the vessels in the chorionic plate, maternal decidua, and trophoblast giant cells. (C) Higher magnification of panel B is given to demonstrate staining in giant cells (arrowheads). (D) VEGF-C was detected in the allantoic mesoderm (arrowheads) and chorion of WT E9.5 placenta by immunostaining but not in allantoic mesoderm of Cyr61 −/− placenta (E). (F) In Northern blot analysis, serum-starved mouse embryonic fibroblasts isolated from E14.5 WT embryos were treated with purified recombinant CYR61 for various durations before the total RNA was isolated. Vegf-C mRNA was upregulated sixfold after 6 h of CYR61 treatment, and this upregulation was sustained for at least 24 h. The hybridization signal was quantified by Phosphorimager. Al, allantois; Ch, chorion; De, decidua; G, giant cells; La, labyrinth; Sp, spongiotrophoblast; U, umbilical cord. Yellow bars, 50 μm.

Techniques Used: Staining, Immunostaining, Northern Blot, Isolation, Purification, Recombinant, Hybridization

14) Product Images from "Soluble vascular endothelial growth factor receptor 3 is essential for corneal alymphaticity"

Article Title: Soluble vascular endothelial growth factor receptor 3 is essential for corneal alymphaticity

Journal: Blood

doi: 10.1182/blood-2012-08-453043

sVEGFR-3 is expressed in corneal epithelium and antagonizes VEGF-C. (A) Reverse-transcriptase PCRs (RT-PCRs) with intron-tail–specific reverse primer and exon-exon junction forward primers showing sVEGFR-3 in mouse cornea. Membrane VEGFR-3 mRNA expression in sclera only. (B) Western blot of corneal and scleral lysate (n = 5) with anti–VEGFR-3 N-terminal antibody demonstrates sVEGFR-3 at 60 kDa in cornea and membrane VEGFR-3 at 170 kDa in sclera. (C) Western blot of corneal and scleral lysate (n = 5) with anti–VEGFR-3 C-terminal antibody demonstrates the expression of membrane VEGFR-3 at 170 kDa in sclera only (none in cornea). (D) Immunolocalization of sVEGFR-3 (brown) in human cornea via an intron-derived C-terminal tail, human sVEGFR-3 antibody (PA 4835; Thermo Scientific). Isotype-negative control rabbit IgG. (E) RT-PCR and western blot of mouse cornea shows VEGF-C mRNA and protein. (F) Western blot of corneal lysate (n = 5) with anti–VEGFR-3 N-terminal antibody, blotted with anti–VEGF-C antibody under reducing (1) and native conditions (2), reveals binding of sVEGFR-3 to VEGF-C. (G) Sandwich ELISA with anti–VEGF-C–coated antibodies in 96-well plates, followed by the addition of corneal lysate (n = 5, each group; corneas from 3 mice in each sample), and then anti–VEGF-C, anti–VEGFR-3 N-terminal, and anti–N-terminal VEGFR-2 antibodies. (H) Competitive ELISA with human recombinant VEGF-C coated on 96-well plates and equimolar human recombinant VEGFR-3 and VEGFR-2 with extracellular domains added, showing affinity and binding to VEGF-C (n = 5). GAPDH, glyceraldehyde-3-phosphate dehydrogenase; IP, immunoprecipitation; M, marker; mVEGFR-3, membrane VEGFR-3; WB, western blot.
Figure Legend Snippet: sVEGFR-3 is expressed in corneal epithelium and antagonizes VEGF-C. (A) Reverse-transcriptase PCRs (RT-PCRs) with intron-tail–specific reverse primer and exon-exon junction forward primers showing sVEGFR-3 in mouse cornea. Membrane VEGFR-3 mRNA expression in sclera only. (B) Western blot of corneal and scleral lysate (n = 5) with anti–VEGFR-3 N-terminal antibody demonstrates sVEGFR-3 at 60 kDa in cornea and membrane VEGFR-3 at 170 kDa in sclera. (C) Western blot of corneal and scleral lysate (n = 5) with anti–VEGFR-3 C-terminal antibody demonstrates the expression of membrane VEGFR-3 at 170 kDa in sclera only (none in cornea). (D) Immunolocalization of sVEGFR-3 (brown) in human cornea via an intron-derived C-terminal tail, human sVEGFR-3 antibody (PA 4835; Thermo Scientific). Isotype-negative control rabbit IgG. (E) RT-PCR and western blot of mouse cornea shows VEGF-C mRNA and protein. (F) Western blot of corneal lysate (n = 5) with anti–VEGFR-3 N-terminal antibody, blotted with anti–VEGF-C antibody under reducing (1) and native conditions (2), reveals binding of sVEGFR-3 to VEGF-C. (G) Sandwich ELISA with anti–VEGF-C–coated antibodies in 96-well plates, followed by the addition of corneal lysate (n = 5, each group; corneas from 3 mice in each sample), and then anti–VEGF-C, anti–VEGFR-3 N-terminal, and anti–N-terminal VEGFR-2 antibodies. (H) Competitive ELISA with human recombinant VEGF-C coated on 96-well plates and equimolar human recombinant VEGFR-3 and VEGFR-2 with extracellular domains added, showing affinity and binding to VEGF-C (n = 5). GAPDH, glyceraldehyde-3-phosphate dehydrogenase; IP, immunoprecipitation; M, marker; mVEGFR-3, membrane VEGFR-3; WB, western blot.

Techniques Used: Expressing, Western Blot, Derivative Assay, Negative Control, Reverse Transcription Polymerase Chain Reaction, Binding Assay, Sandwich ELISA, Mouse Assay, Competitive ELISA, Recombinant, Immunoprecipitation, Marker

Corneal injury induces lymphangiogenesis, upregulation of VEGF-C, and membrane VEGFR-3 expression. (A) Immunostaining of normal and sutured cornea on day 3 reveals sVEGFR-3 and VEGF-C upregulation at the site of suture, indicating that sVEGFR-3 initially tends to capture VEGF-C and control VEGF-C surge. Negative controls are sections stained with isotype-control primary antibodies. (B) Real-time PCR of normal and sutured cornea on day 3 revealing mechanical trauma leads to instant increase in VEGF-C mRNA levels (n = 5). (C) Western blot of normal (1) and sutured cornea (2) on day 5 leads to expression of membrane VEGFR-3 (170 kDa) in sutured cornea that leads to signaling through VEGFR-3 and development of lymphatic vessels.
Figure Legend Snippet: Corneal injury induces lymphangiogenesis, upregulation of VEGF-C, and membrane VEGFR-3 expression. (A) Immunostaining of normal and sutured cornea on day 3 reveals sVEGFR-3 and VEGF-C upregulation at the site of suture, indicating that sVEGFR-3 initially tends to capture VEGF-C and control VEGF-C surge. Negative controls are sections stained with isotype-control primary antibodies. (B) Real-time PCR of normal and sutured cornea on day 3 revealing mechanical trauma leads to instant increase in VEGF-C mRNA levels (n = 5). (C) Western blot of normal (1) and sutured cornea (2) on day 5 leads to expression of membrane VEGFR-3 (170 kDa) in sutured cornea that leads to signaling through VEGFR-3 and development of lymphatic vessels.

Techniques Used: Expressing, Immunostaining, Staining, Real-time Polymerase Chain Reaction, Western Blot

sVEGFR-3 knockdown leads to growth of corneal lymphatic and blood vessels and expression and phosphorylation of membrane VEGFR-3. (A) Immunofluorescent staining and confocal imaging of cornea flat mounts (n = 10 each group) injected with 2 μg pshRNA–sVEGFR-3, pshRNA-CTR, pshRNA–sVEGFR-3 + anti–VEGF-C immunoglobulin, and IgG. pshRNA–sVEGFR-3 injection leads to lymphangiogenesis in 10 days, with vessels oriented to injection site. Anti–VEGF-C antibody administration 1 day before pshRNA-sVEGFR-3 controls lymphangiogenesis. (B-C) Corneal area of blood and lymphatic vessels 10 days after pshRNA–sVEGFR-3, pshRNA-CTR, pshRNA–sVEGFR-3 + anti–VEGF-C immunoglobulin, and IgG antibody injection. Anti–VEGF-C immunoglobulin injection 1 day prior to pshRNA–sVEGFR-3 shows 75% and 56% reduction of lymphatic and blood vessels (n = 10 each group). (D) sVEGFR-3 knockdown with pshRNA–sVEGFR-3 injection into cornea is associated with expression of membrane VEGFR-3 and downregulation of sVEGFR-3 as seen in western blot with anti–N-terminal VEGFR-3 antibody. (E) sVEGFR-3 knockdown with pshRNA–sVEGFR-3 (samples harvested on day 5 after injection) is associated with phosphorylation of membrane VEGFR-3 as demonstrated via immunoprecipitation with anti–N-terminal VEGFR-3 antibody followed by western blotting with anti-phosphotyrosine antibody. (F) Stripping and reblotting the membrane with anti–N-terminal end VEGFR-3 antibody confirms that sVEGFR-3 is present after control shRNA treatment but not after pshRNA–sVEGFR-3. * P
Figure Legend Snippet: sVEGFR-3 knockdown leads to growth of corneal lymphatic and blood vessels and expression and phosphorylation of membrane VEGFR-3. (A) Immunofluorescent staining and confocal imaging of cornea flat mounts (n = 10 each group) injected with 2 μg pshRNA–sVEGFR-3, pshRNA-CTR, pshRNA–sVEGFR-3 + anti–VEGF-C immunoglobulin, and IgG. pshRNA–sVEGFR-3 injection leads to lymphangiogenesis in 10 days, with vessels oriented to injection site. Anti–VEGF-C antibody administration 1 day before pshRNA-sVEGFR-3 controls lymphangiogenesis. (B-C) Corneal area of blood and lymphatic vessels 10 days after pshRNA–sVEGFR-3, pshRNA-CTR, pshRNA–sVEGFR-3 + anti–VEGF-C immunoglobulin, and IgG antibody injection. Anti–VEGF-C immunoglobulin injection 1 day prior to pshRNA–sVEGFR-3 shows 75% and 56% reduction of lymphatic and blood vessels (n = 10 each group). (D) sVEGFR-3 knockdown with pshRNA–sVEGFR-3 injection into cornea is associated with expression of membrane VEGFR-3 and downregulation of sVEGFR-3 as seen in western blot with anti–N-terminal VEGFR-3 antibody. (E) sVEGFR-3 knockdown with pshRNA–sVEGFR-3 (samples harvested on day 5 after injection) is associated with phosphorylation of membrane VEGFR-3 as demonstrated via immunoprecipitation with anti–N-terminal VEGFR-3 antibody followed by western blotting with anti-phosphotyrosine antibody. (F) Stripping and reblotting the membrane with anti–N-terminal end VEGFR-3 antibody confirms that sVEGFR-3 is present after control shRNA treatment but not after pshRNA–sVEGFR-3. * P

Techniques Used: Expressing, Staining, Imaging, Injection, Western Blot, Immunoprecipitation, Stripping Membranes, shRNA

sVEGFR-3 inhibits VEGF-C–induced VEGFR-2 phosphorylation but not VEGF-A–induced VEGFR-2 phosphorylation. (A) Immunoprecipitation by VEGFR-2 antibody from HUVEC lysate. The input shows soluble and membrane VEGFR-3 bands. However, VEGFR-3 bands were not detected after precipitation, indicating that the two receptors do not heterodimerize. (B) After 24 hours of serum starvation, HUVEC was stimulated with 20 ng/mL VEGF-A or 20 ng/mL VEGF-A + 500 ng/mL recombinant sVEGFR-3. Recombinant sVEGFR-3 did not block VEGF-A–induced VEGFR-2 phosphorylation. (C) After 24 hours of serum starvation, HUVEC was stimulated with 20 ng/mL, 100 ng/mL, or 500 ng/mL VEGF-C or 500 ng/mL VEGF-C + 1750 ng/mL recombinant sVEGFR-3. Recombinant sVEGFR-3 treatment inhibits VEGF-C–induced VEGFR-2 phosphorylation. IP, immunoprecipitation; WB, western blot.
Figure Legend Snippet: sVEGFR-3 inhibits VEGF-C–induced VEGFR-2 phosphorylation but not VEGF-A–induced VEGFR-2 phosphorylation. (A) Immunoprecipitation by VEGFR-2 antibody from HUVEC lysate. The input shows soluble and membrane VEGFR-3 bands. However, VEGFR-3 bands were not detected after precipitation, indicating that the two receptors do not heterodimerize. (B) After 24 hours of serum starvation, HUVEC was stimulated with 20 ng/mL VEGF-A or 20 ng/mL VEGF-A + 500 ng/mL recombinant sVEGFR-3. Recombinant sVEGFR-3 did not block VEGF-A–induced VEGFR-2 phosphorylation. (C) After 24 hours of serum starvation, HUVEC was stimulated with 20 ng/mL, 100 ng/mL, or 500 ng/mL VEGF-C or 500 ng/mL VEGF-C + 1750 ng/mL recombinant sVEGFR-3. Recombinant sVEGFR-3 treatment inhibits VEGF-C–induced VEGFR-2 phosphorylation. IP, immunoprecipitation; WB, western blot.

Techniques Used: Immunoprecipitation, Recombinant, Blocking Assay, Western Blot

15) Product Images from "VEGFR3 Inhibition Chemosensitizes Ovarian Cancer Stemlike Cells through Down-Regulation of BRCA1 and BRCA2"

Article Title: VEGFR3 Inhibition Chemosensitizes Ovarian Cancer Stemlike Cells through Down-Regulation of BRCA1 and BRCA2

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

doi: 10.1016/j.neo.2014.04.003

Expression of VEGF-C, VEGF-D, and VEGFR3 in ovarian cancer. (A and B) qRT-PCR evaluation of VEGF-C mRNA and VEGFR3 mRNA in VLCs, TECs, and primary ovarian tumor cells (TCs) is presented. (C) Western blot confirms high VEGF-C protein expression in VLCs.
Figure Legend Snippet: Expression of VEGF-C, VEGF-D, and VEGFR3 in ovarian cancer. (A and B) qRT-PCR evaluation of VEGF-C mRNA and VEGFR3 mRNA in VLCs, TECs, and primary ovarian tumor cells (TCs) is presented. (C) Western blot confirms high VEGF-C protein expression in VLCs.

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot

VEGFR3 inhibition decreases BRCA1 and BRCA2 gene expression through p-ERK and E2F1. (A) (i) Phosphoprotein Western blot analysis of control and VEGF-C–treated A2780 and OVCAR8 cells in the presence or absence of Maz51 showing VEGF-C treatment
Figure Legend Snippet: VEGFR3 inhibition decreases BRCA1 and BRCA2 gene expression through p-ERK and E2F1. (A) (i) Phosphoprotein Western blot analysis of control and VEGF-C–treated A2780 and OVCAR8 cells in the presence or absence of Maz51 showing VEGF-C treatment

Techniques Used: Inhibition, Expressing, Western Blot

16) Product Images from "Fluid shear stress regulates vascular remodeling via VEGFR-3 activation, although independently of its ligand, VEGF-C, in the uterus during pregnancy"

Article Title: Fluid shear stress regulates vascular remodeling via VEGFR-3 activation, although independently of its ligand, VEGF-C, in the uterus during pregnancy

Journal: International Journal of Molecular Medicine

doi: 10.3892/ijmm.2017.3108

Vascular endothelial growth factor receptor-3 (VEGFR-3) expression in the endometrium does not coincide with that of lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1) and is independent of VEGF-C. (A) Image showing LYVE-1 + regions and VEGFR-3 + regions in the uterus at 6.5 days post coitum (dpc). Scale bars, 500 µ m. (B) Image showing LYVE-1 + regions and VEGF-C + regions in the uterus at 6.5 dpc. Scale bars, 500 µ m. (C) Magnified images showing VEGFR-3 + regions and LYVE-1 + regions in the endometrium and myometrium of uterus at 6.5 dpc. Scale bars, 50 µ m. (D) Magnified images showing VEGF-C + regions and LYVE-1 + regions in the endometrium and myometrium of uterus at 6.5 dpc. Scale bars, 50 µ m. (E) Comparison of VEGF-C protein levels in the uterus of 4.5–8.5 dpc. Levels of VEGF-C were normalized to those of β-actin. MR, mesometrial region; AMR, anti-mesometrial region; UL, uterus lumen; VSR, venous sinus region; Em, embryo.
Figure Legend Snippet: Vascular endothelial growth factor receptor-3 (VEGFR-3) expression in the endometrium does not coincide with that of lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1) and is independent of VEGF-C. (A) Image showing LYVE-1 + regions and VEGFR-3 + regions in the uterus at 6.5 days post coitum (dpc). Scale bars, 500 µ m. (B) Image showing LYVE-1 + regions and VEGF-C + regions in the uterus at 6.5 dpc. Scale bars, 500 µ m. (C) Magnified images showing VEGFR-3 + regions and LYVE-1 + regions in the endometrium and myometrium of uterus at 6.5 dpc. Scale bars, 50 µ m. (D) Magnified images showing VEGF-C + regions and LYVE-1 + regions in the endometrium and myometrium of uterus at 6.5 dpc. Scale bars, 50 µ m. (E) Comparison of VEGF-C protein levels in the uterus of 4.5–8.5 dpc. Levels of VEGF-C were normalized to those of β-actin. MR, mesometrial region; AMR, anti-mesometrial region; UL, uterus lumen; VSR, venous sinus region; Em, embryo.

Techniques Used: Expressing

17) Product Images from "Recombinant canstatin inhibits VEGF‐A‐induced lymphangiogenesis and metastasis in an oral squamous cell carcinoma SCC‐VII animal model"

Article Title: Recombinant canstatin inhibits VEGF‐A‐induced lymphangiogenesis and metastasis in an oral squamous cell carcinoma SCC‐VII animal model

Journal: Cancer Medicine

doi: 10.1002/cam4.866

Effects of recombinant canstatin on the expression of vascular endothelial growth factor ( VEGF ) family proteins in CoCl 2 ‐treated squamous cell carcinoma ( SCC )‐ VII cells. (A) SCC ‐ VII cells were treated with different concentrations of recombinant canstatin (0, 0.5, 40 μ g/mL) in the presence of 100 μ mol/L CoCl 2 , and incubated for 24 h. cDNA s were generated from DN ase I‐treated total RNA , and PCR was performed with specific primers for vascular endothelial growth factor ( VEGF )‐A, ‐B, ‐C, and β ‐actin. (B) The PCR products from three independent experiments in (A) were quantified and are represented as a bar diagram. The transcript levels of VEGF ‐A, ‐B, and ‐C mRNA in the control (recombinant canstatin‐ and CoCl 2 ‐untreated cells) were established as 100%. (C) Protein levels of VEGF ‐A and ‐C in the intracellular fraction were determined using western blot analysis with anti‐ VEGF ‐A and anti‐ VEGF ‐C antibodies. (D) The amounts of VEGF ‐A and ‐C obtained in three independent experiments of (C) were quantified and are represented as a bar diagram. The levels of VEGF ‐A and ‐C in the control were established as 100%. Data are presented as mean ± SD of three independent experiments (* P
Figure Legend Snippet: Effects of recombinant canstatin on the expression of vascular endothelial growth factor ( VEGF ) family proteins in CoCl 2 ‐treated squamous cell carcinoma ( SCC )‐ VII cells. (A) SCC ‐ VII cells were treated with different concentrations of recombinant canstatin (0, 0.5, 40 μ g/mL) in the presence of 100 μ mol/L CoCl 2 , and incubated for 24 h. cDNA s were generated from DN ase I‐treated total RNA , and PCR was performed with specific primers for vascular endothelial growth factor ( VEGF )‐A, ‐B, ‐C, and β ‐actin. (B) The PCR products from three independent experiments in (A) were quantified and are represented as a bar diagram. The transcript levels of VEGF ‐A, ‐B, and ‐C mRNA in the control (recombinant canstatin‐ and CoCl 2 ‐untreated cells) were established as 100%. (C) Protein levels of VEGF ‐A and ‐C in the intracellular fraction were determined using western blot analysis with anti‐ VEGF ‐A and anti‐ VEGF ‐C antibodies. (D) The amounts of VEGF ‐A and ‐C obtained in three independent experiments of (C) were quantified and are represented as a bar diagram. The levels of VEGF ‐A and ‐C in the control were established as 100%. Data are presented as mean ± SD of three independent experiments (* P

Techniques Used: Recombinant, Expressing, Incubation, Generated, Polymerase Chain Reaction, Western Blot

Effects of recombinant canstatin on the expression of vascular endothelial growth factor ( VEGF )‐A, VEGF ‐C, vascular endothelial growth factor receptors ( VEGFR )‐1, VEGFR ‐2, and VEGFR ‐3 in squamous cell carcinoma ( SCC )‐ VII ‐induced tumors. (A, C) The presence of VEGF ‐A, VEGF ‐C (A), VEGFR ‐1, VEGFR ‐2, and VEGFR ‐3 (C) in the sections of PBS ‐treated or 5 mg/kg/day recombinant canstatin‐treated SCC ‐ VII ‐induced tumors was determined using immunohistochemical analysis. All tumor sections were digitized and images were captured under a 400× objective magnification. Scale bar = 100 μ m. (B, D) Immunohistochemical intensities of VEGF ‐A, VEGF ‐C (B), VEGFR ‐1, VEGFR ‐2, and VEGFR ‐3 (D) from captured images were analyzed using the Image J program and are represented as bar diagrams. The intensities of the VEGF ‐A, VEGF ‐C, VEGFR ‐1, VEGFR ‐2, and VEGFR ‐3 staining of the PBS ‐treated control were established as 100%. Data are presented as mean ± SD of three independent experiments (*** P
Figure Legend Snippet: Effects of recombinant canstatin on the expression of vascular endothelial growth factor ( VEGF )‐A, VEGF ‐C, vascular endothelial growth factor receptors ( VEGFR )‐1, VEGFR ‐2, and VEGFR ‐3 in squamous cell carcinoma ( SCC )‐ VII ‐induced tumors. (A, C) The presence of VEGF ‐A, VEGF ‐C (A), VEGFR ‐1, VEGFR ‐2, and VEGFR ‐3 (C) in the sections of PBS ‐treated or 5 mg/kg/day recombinant canstatin‐treated SCC ‐ VII ‐induced tumors was determined using immunohistochemical analysis. All tumor sections were digitized and images were captured under a 400× objective magnification. Scale bar = 100 μ m. (B, D) Immunohistochemical intensities of VEGF ‐A, VEGF ‐C (B), VEGFR ‐1, VEGFR ‐2, and VEGFR ‐3 (D) from captured images were analyzed using the Image J program and are represented as bar diagrams. The intensities of the VEGF ‐A, VEGF ‐C, VEGFR ‐1, VEGFR ‐2, and VEGFR ‐3 staining of the PBS ‐treated control were established as 100%. Data are presented as mean ± SD of three independent experiments (*** P

Techniques Used: Recombinant, Expressing, Immunohistochemistry, Staining

Related Articles

Incubation:

Article Title: Grafting and Early Expression of Growth Factors from Adipose-Derived Stem Cells Transplanted into the Cochlea, in a Guinea Pig Model of Acoustic Trauma
Article Snippet: .. The specimens were then incubated overnight at 4°C with a solution containing rabbit polyclonal anti-VEGF-A (diluted 1:100, Bioss Woburn, MA, USA), anti-VEGF-C (diluted 1:50, Santa Cruz Dallas, TX, USA), anti-PDGFR (diluted 1:500, Abcam Cambridge, UK), and anti-TGFβ1 (diluted 1:100, Antibodies, Atlanta, GA, USA) primary antibodies. .. All slides were then washed twice in PBS and incubated at room temperature for 2 h, light-protected, with labeled conjugated goat anti-rabbit secondary antibody (Alexa Fluor 633, IgG; Invitrogen, Carlsbad, CA, USA) diluted 1:400 in 0.1M PBS.

Article Title: Modulation the crosstalk between tumor-associated macrophages and non-small cell lung cancer to inhibit tumor migration and invasion by ginsenoside Rh2
Article Snippet: .. After deparaffinization, endogenous peroxidase activity was blocked by incubation with 3% peroxide-methanol solution at room temperature (RT) for 10 min, and then antigen retrieval was performed at 100 °C in an autoclave for 7 min. After washing with PBS, sections were incubated with primary antibodies against theCD206 monoclonal antibody (clone 10D6, Zhongshan Goldenbridge Biotechnology Co., LTD., Beijing, China) and VEGF-C (Santa Cruz Biotechnology, Santa Cruz, CA, USA)overnight at 4 °C. .. Next, sections were incubated with aDAKO EnVision kit (DAKO, Glostrup, Denmark) following the manufacturer’s instructions.

other:

Article Title: MT1-MMP sheds LYVE-1 on lymphatic endothelial cells and suppresses VEGF-C production to inhibit lymphangiogenesis
Article Snippet: AntibodiesThe antibodies used in this study include the following: anti-CD31 antibody (MEC13.3, 553070, BD Pharmingen; 1:150); anti-LYVE-1 antibody (11-034, AngioBio; 1:250); anti-VEGFR3 antibody (AF743, R & D System; 1:200); anti-MT1-MMP antibody (ab51074, Abcam; 1:2,000); anti-CD11b antibody (5573-94, BD Pharmingen; 1:200); an Alexa Fluor 488-conjugated anti-F4/80 antibody (MCA497A488, AbD Secrotec; 1:150); anti-BrdU antibody (Bu20a, M0744, Dako; 1:250); anti-VEGF-C (H-190, Santa Cruz, 1:1,000); anti-p65 (H-286, Santa Cruz; 1:2,000); anti-Ikβα (C-21, sc-371, Santa Cruz, 1:2,000); anti-Ikββ (S-20, sc-946, Santa Cruz, 1:2,000); anti-Akt (9272, Cell Signaling, 1:2,000); anti-p-Akt (Ser473) (9271, Cell Signaling, 1:2,000); β-actin (A1978, Sigma, 1:8,000); fluorescein isothiocyanate-conjugated goat anti-rabbit antibody (4050-02, Southern Biotech; 1:500); fluorescein isothiocyanate-conjugated donkey anti-rat antibody (6430-02, Southern Biotech; 1:500); Alexa Fluor 594-conjugated goat anti-rabbit antibody (A-11012, Invitrogen, 1:600); and Alexa Fluor 594-conjugated donkey anti-goat antibody (A11058, Invitrogen; 1:500).

Activity Assay:

Article Title: Modulation the crosstalk between tumor-associated macrophages and non-small cell lung cancer to inhibit tumor migration and invasion by ginsenoside Rh2
Article Snippet: .. After deparaffinization, endogenous peroxidase activity was blocked by incubation with 3% peroxide-methanol solution at room temperature (RT) for 10 min, and then antigen retrieval was performed at 100 °C in an autoclave for 7 min. After washing with PBS, sections were incubated with primary antibodies against theCD206 monoclonal antibody (clone 10D6, Zhongshan Goldenbridge Biotechnology Co., LTD., Beijing, China) and VEGF-C (Santa Cruz Biotechnology, Santa Cruz, CA, USA)overnight at 4 °C. .. Next, sections were incubated with aDAKO EnVision kit (DAKO, Glostrup, Denmark) following the manufacturer’s instructions.

Western Blot:

Article Title: Therapeutic Impact of Nanoparticle Therapy Targeting Tumor Associate Macrophages
Article Snippet: .. Anti-VEGF-C (Santa Cruz, sc-1881) was used at a dilution of 1:200 for western blot analysis. .. Single cell suspensions from treated tumors were analyzed via FACS using anti-GRP78 as previously described ( ).

Article Title: Overexpression of both VEGF-A and VEGF-C in gastric cancer correlates with prognosis, and silencing of both is effective to inhibit cancer growth
Article Snippet: .. Western blotting was done using antibodies against VEGF-A and VEGF-C (Santa Cruz Biotechnology, CA, USA). .. All the bands were visualized with the enhanced chemiluminescence (Pierce, Rockford, IL).

Staining:

Article Title: Therapeutic Lymphangiogenesis With Implantation of Adipose-Derived Regenerative Cells
Article Snippet: .. Frozen sections were stained with anti–VEGF‐C (Santa Cruz Biotechnology, Inc), anti–LYVE‐1 (Acris), anti‐CD11b (SRT AbD), or anti‐CD163 (Santa Cruz Biotechnology, Inc), and the nuclei were stained with 4′,6‐diamidino‐2‐phenylindole (DAPI). ..

Recombinant:

Article Title: Production of Vascular Endothelial Growth Factors from Human Lung Macrophages Induced by Group IIA and Group X Secreted Phospholipases A2
Article Snippet: .. The following were purchased: LPS (from Escherichia coli serotype 026:B6), 5′-(N-ethylcarboxamido)-adenosine (NECA), 2- p -(2-Carboxyethyl)phenethylamino-5′-N-ethylcarboxamidoadenosine , 2-Chloro-N6 -(3-iodobenzyl)-adenosine-5′-N-methyluronamide (Cl-IB-MECA), Pipes, BSA, Percoll, l -glutamine, antibiotic-antimycotic solution (10,000 UI/ml penicillin, 10 mg/ml streptomycin, and 25 μg/ml amphotericin B), Triton X-100, Polymyxin B (Sigma-Aldrich, St. Louis, MO); RPMI 1640, FCS, and guanidine hydrochloride (MP Biomedicals Europe, Illkirch, France); rabbit polyclonal anti–VEGF-B (H-70), anti–VEGF-C (H-190), and anti–VEGF-D (H-144) Abs, goat polyclonal anti-PlGF (C-20) and anti-GAPDH (V-18) Abs, normal rabbit and goat IgG Abs, HRP-conjugated donkey anti-rabbit and anti-goat IgG Abs (Santa Cruz Biotechnology, Santa Cruz, CA); goat polyclonal anti–VEGF-A Ab, human recombinant VEGF-A165 , and VEGF-A121 (R & D System, Minneapolis, MN); SB203580 (Calbiochem, La Jolla, CA); PD98059 (Cell Signaling, Beverly, MA). .. The sPLA2 inhibitors Me-Indoxam and RO092906A ( , ), recombinant hGIIA and hGX sPLA2 s , and the H48Q mutants of hGIIA and hGX (hGIIA-H48Q and hGX-H48Q) ( ) were prepared as described.

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    Santa Cruz Biotechnology anti vegf c
    sPLA 2 s induce the release of <t>VEGF-A</t> and <t>VEGF-C</t> from HLMs. A and B , HLMs were incubated (37°C, 24 h) with RPMI 1640 alone (Control) or with the indicated concentrations of hGIIA, hGX, or LPS. VEGF-A ( A ) and VEGF-C ( B ) release was determined by ELISA. Data are mean ± SEM of four experiments. * p
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    sPLA 2 s induce the release of VEGF-A and VEGF-C from HLMs. A and B , HLMs were incubated (37°C, 24 h) with RPMI 1640 alone (Control) or with the indicated concentrations of hGIIA, hGX, or LPS. VEGF-A ( A ) and VEGF-C ( B ) release was determined by ELISA. Data are mean ± SEM of four experiments. * p

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: Production of Vascular Endothelial Growth Factors from Human Lung Macrophages Induced by Group IIA and Group X Secreted Phospholipases A2

    doi: 10.4049/jimmunol.0902501

    Figure Lengend Snippet: sPLA 2 s induce the release of VEGF-A and VEGF-C from HLMs. A and B , HLMs were incubated (37°C, 24 h) with RPMI 1640 alone (Control) or with the indicated concentrations of hGIIA, hGX, or LPS. VEGF-A ( A ) and VEGF-C ( B ) release was determined by ELISA. Data are mean ± SEM of four experiments. * p

    Article Snippet: The following were purchased: LPS (from Escherichia coli serotype 026:B6), 5′-(N-ethylcarboxamido)-adenosine (NECA), 2- p -(2-Carboxyethyl)phenethylamino-5′-N-ethylcarboxamidoadenosine , 2-Chloro-N6 -(3-iodobenzyl)-adenosine-5′-N-methyluronamide (Cl-IB-MECA), Pipes, BSA, Percoll, l -glutamine, antibiotic-antimycotic solution (10,000 UI/ml penicillin, 10 mg/ml streptomycin, and 25 μg/ml amphotericin B), Triton X-100, Polymyxin B (Sigma-Aldrich, St. Louis, MO); RPMI 1640, FCS, and guanidine hydrochloride (MP Biomedicals Europe, Illkirch, France); rabbit polyclonal anti–VEGF-B (H-70), anti–VEGF-C (H-190), and anti–VEGF-D (H-144) Abs, goat polyclonal anti-PlGF (C-20) and anti-GAPDH (V-18) Abs, normal rabbit and goat IgG Abs, HRP-conjugated donkey anti-rabbit and anti-goat IgG Abs (Santa Cruz Biotechnology, Santa Cruz, CA); goat polyclonal anti–VEGF-A Ab, human recombinant VEGF-A165 , and VEGF-A121 (R & D System, Minneapolis, MN); SB203580 (Calbiochem, La Jolla, CA); PD98059 (Cell Signaling, Beverly, MA).

    Techniques: Incubation, Enzyme-linked Immunosorbent Assay

    HLMs constitutively express different forms of VEGF. A , Expression of VEGF mRNAs. RNA extraction from resting HLMs and RT-PCR was performed as described under Materials and Methods . Specific RT-PCR amplification products for VEGFA (isoforms 189, 165, and 121), VEGFB (isoforms 186 and 167), VEGFC, VEGFD, PlGF , and GAPDH were separated on 2% agarose gel, stained with ethidium bromide, and visualized with an image analysis system. The experiment shown is representative of three separate experiments. B , Detection of VEGF proteins. HLM protein extracts (40 μg per sample) were immunoblotted with anti–VEGF-A (gel I and II), anti-PlGF (gel III), anti–VEGF-B (gel IV), anti–VEGF-C (gel V), and anti–VEGF-D (gel VI) Abs. rhVEGF-A 165 , MCF-7 cells, EBNA expressing PlGF-1, RAW 264.7 cells were used as positive controls. Stripped membranes were reprobed with anti-GAPDH Ab to confirm equal protein content of each sample. Each Western blot shown is representative of three separate experiments.

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: Production of Vascular Endothelial Growth Factors from Human Lung Macrophages Induced by Group IIA and Group X Secreted Phospholipases A2

    doi: 10.4049/jimmunol.0902501

    Figure Lengend Snippet: HLMs constitutively express different forms of VEGF. A , Expression of VEGF mRNAs. RNA extraction from resting HLMs and RT-PCR was performed as described under Materials and Methods . Specific RT-PCR amplification products for VEGFA (isoforms 189, 165, and 121), VEGFB (isoforms 186 and 167), VEGFC, VEGFD, PlGF , and GAPDH were separated on 2% agarose gel, stained with ethidium bromide, and visualized with an image analysis system. The experiment shown is representative of three separate experiments. B , Detection of VEGF proteins. HLM protein extracts (40 μg per sample) were immunoblotted with anti–VEGF-A (gel I and II), anti-PlGF (gel III), anti–VEGF-B (gel IV), anti–VEGF-C (gel V), and anti–VEGF-D (gel VI) Abs. rhVEGF-A 165 , MCF-7 cells, EBNA expressing PlGF-1, RAW 264.7 cells were used as positive controls. Stripped membranes were reprobed with anti-GAPDH Ab to confirm equal protein content of each sample. Each Western blot shown is representative of three separate experiments.

    Article Snippet: The following were purchased: LPS (from Escherichia coli serotype 026:B6), 5′-(N-ethylcarboxamido)-adenosine (NECA), 2- p -(2-Carboxyethyl)phenethylamino-5′-N-ethylcarboxamidoadenosine , 2-Chloro-N6 -(3-iodobenzyl)-adenosine-5′-N-methyluronamide (Cl-IB-MECA), Pipes, BSA, Percoll, l -glutamine, antibiotic-antimycotic solution (10,000 UI/ml penicillin, 10 mg/ml streptomycin, and 25 μg/ml amphotericin B), Triton X-100, Polymyxin B (Sigma-Aldrich, St. Louis, MO); RPMI 1640, FCS, and guanidine hydrochloride (MP Biomedicals Europe, Illkirch, France); rabbit polyclonal anti–VEGF-B (H-70), anti–VEGF-C (H-190), and anti–VEGF-D (H-144) Abs, goat polyclonal anti-PlGF (C-20) and anti-GAPDH (V-18) Abs, normal rabbit and goat IgG Abs, HRP-conjugated donkey anti-rabbit and anti-goat IgG Abs (Santa Cruz Biotechnology, Santa Cruz, CA); goat polyclonal anti–VEGF-A Ab, human recombinant VEGF-A165 , and VEGF-A121 (R & D System, Minneapolis, MN); SB203580 (Calbiochem, La Jolla, CA); PD98059 (Cell Signaling, Beverly, MA).

    Techniques: Expressing, RNA Extraction, Reverse Transcription Polymerase Chain Reaction, Amplification, Agarose Gel Electrophoresis, Staining, Western Blot

    Immunohistochemical results for blood vessel remodeling in acute colitis ( A ). Comparison of microvessel density between AD-VEGF-C treated and DSS-treated mice ( B ) and between AD-VEGF-C156S-treated and PBS-treated mice ( C ). Both AD-VEGF-C and VEGF-C156S induced a significant increase in microvessel density compared to DSS-treated mice (both P

    Journal: Brazilian Journal of Medical and Biological Research

    Article Title: Promoting inflammatory lymphangiogenesis by vascular endothelial growth factor-C (VEGF-C) aggravated intestinal inflammation in mice with experimental acute colitis

    doi: 10.1590/1414-431X20154738

    Figure Lengend Snippet: Immunohistochemical results for blood vessel remodeling in acute colitis ( A ). Comparison of microvessel density between AD-VEGF-C treated and DSS-treated mice ( B ) and between AD-VEGF-C156S-treated and PBS-treated mice ( C ). Both AD-VEGF-C and VEGF-C156S induced a significant increase in microvessel density compared to DSS-treated mice (both P

    Article Snippet: One limitation of the present study was that, due to the lack of a commercially available VEGF-C antibody, which can be used on murine specimens for confocal microscopy, we could not determine the identity of the cells that produced VEGF-C. Further studies are necessary to understand the role and functions of lymphangiogenesis in IBD.

    Techniques: Immunohistochemistry, Mouse Assay

    A , Western blot of VEGF-C, VEGFR-2 and VEGFR-3 in the different experimental groups. B , Comparison of VEGFR-3 mRNA expression detected by quantitative real-time RT-PCR. VEGFR-3 mRNA levels were significantly upregulated in VEGF-C156S-treated mice compared to DSS-treated mice (P

    Journal: Brazilian Journal of Medical and Biological Research

    Article Title: Promoting inflammatory lymphangiogenesis by vascular endothelial growth factor-C (VEGF-C) aggravated intestinal inflammation in mice with experimental acute colitis

    doi: 10.1590/1414-431X20154738

    Figure Lengend Snippet: A , Western blot of VEGF-C, VEGFR-2 and VEGFR-3 in the different experimental groups. B , Comparison of VEGFR-3 mRNA expression detected by quantitative real-time RT-PCR. VEGFR-3 mRNA levels were significantly upregulated in VEGF-C156S-treated mice compared to DSS-treated mice (P

    Article Snippet: One limitation of the present study was that, due to the lack of a commercially available VEGF-C antibody, which can be used on murine specimens for confocal microscopy, we could not determine the identity of the cells that produced VEGF-C. Further studies are necessary to understand the role and functions of lymphangiogenesis in IBD.

    Techniques: Western Blot, Expressing, Quantitative RT-PCR, Mouse Assay

    A , Assessment of histological scores in the different groups. Histology of normal tissue is shown in panels a and d , acute colitis was induced with DSS in DSS-treated mice (panels b and e ); in AD-VEGF-C-treated mice (panels c and f ); in PBS-treated mice (panels g and i ); and in VEGF-C156S-treated mice (panels h and j ). DSS-treated mice showed significantly greater histological damage (cellular infiltration, goblet cell depletion, damage to crypt architecture and submucosal edema) ( A: b , e , g , i ) compared to normal mice. VEGF-C-treated mice exposed to DSS showed significantly higher histological scores with more severe histological damage compared to control DSS-treated mice, both in the AD-VEGF-C ( B , P

    Journal: Brazilian Journal of Medical and Biological Research

    Article Title: Promoting inflammatory lymphangiogenesis by vascular endothelial growth factor-C (VEGF-C) aggravated intestinal inflammation in mice with experimental acute colitis

    doi: 10.1590/1414-431X20154738

    Figure Lengend Snippet: A , Assessment of histological scores in the different groups. Histology of normal tissue is shown in panels a and d , acute colitis was induced with DSS in DSS-treated mice (panels b and e ); in AD-VEGF-C-treated mice (panels c and f ); in PBS-treated mice (panels g and i ); and in VEGF-C156S-treated mice (panels h and j ). DSS-treated mice showed significantly greater histological damage (cellular infiltration, goblet cell depletion, damage to crypt architecture and submucosal edema) ( A: b , e , g , i ) compared to normal mice. VEGF-C-treated mice exposed to DSS showed significantly higher histological scores with more severe histological damage compared to control DSS-treated mice, both in the AD-VEGF-C ( B , P

    Article Snippet: One limitation of the present study was that, due to the lack of a commercially available VEGF-C antibody, which can be used on murine specimens for confocal microscopy, we could not determine the identity of the cells that produced VEGF-C. Further studies are necessary to understand the role and functions of lymphangiogenesis in IBD.

    Techniques: Mouse Assay

    Immunohistochemical results for lymphatic remodeling in acute colitis ( A ). Comparison of lymphatic vessel density (LVD) between AD-VEGF-C treated and control mice ( B ) and between AD-VEGF-C156S-treated and control mice ( C ). AD-VEGF-C did not induce an increase in LVD (P=0.50), whereas VEGF-C156S induced a 1.3-fold increase in LVD compared to PBS-treated mice. Comparisons of lymphatic vessel size between AD-VEGF-C-treated and DSS-treated mice are shown in panel D and between AD-VEGF-C156S-treated and PBS-treated mice are shown in panel E . Both AD-VEGF-C and VEGF-C156S-treated mice had significantly greater lymphatic vessel size compared to control mice (both P

    Journal: Brazilian Journal of Medical and Biological Research

    Article Title: Promoting inflammatory lymphangiogenesis by vascular endothelial growth factor-C (VEGF-C) aggravated intestinal inflammation in mice with experimental acute colitis

    doi: 10.1590/1414-431X20154738

    Figure Lengend Snippet: Immunohistochemical results for lymphatic remodeling in acute colitis ( A ). Comparison of lymphatic vessel density (LVD) between AD-VEGF-C treated and control mice ( B ) and between AD-VEGF-C156S-treated and control mice ( C ). AD-VEGF-C did not induce an increase in LVD (P=0.50), whereas VEGF-C156S induced a 1.3-fold increase in LVD compared to PBS-treated mice. Comparisons of lymphatic vessel size between AD-VEGF-C-treated and DSS-treated mice are shown in panel D and between AD-VEGF-C156S-treated and PBS-treated mice are shown in panel E . Both AD-VEGF-C and VEGF-C156S-treated mice had significantly greater lymphatic vessel size compared to control mice (both P

    Article Snippet: One limitation of the present study was that, due to the lack of a commercially available VEGF-C antibody, which can be used on murine specimens for confocal microscopy, we could not determine the identity of the cells that produced VEGF-C. Further studies are necessary to understand the role and functions of lymphangiogenesis in IBD.

    Techniques: Immunohistochemistry, Mouse Assay

    Evaluation of disease activity index (DAI) in AD-VEGF-C-treated mice. Mean DAI scores (±SD) were higher in the AD-VEGF-C-treated mice ( A ) and in recombinant VEGF-C156S-treated mice ( B ), compared to DSS-treated mice. The difference in DAI scores between AD-VEGF-C-treated mice and DSS-treated mice became significant on the second day of observation and continued to differ thereafter. There was a significant difference in DAI scores between recombinant VEGF-C156S-treated mice and PBS-treated mice on days 3, 5, 6 and 7. *P

    Journal: Brazilian Journal of Medical and Biological Research

    Article Title: Promoting inflammatory lymphangiogenesis by vascular endothelial growth factor-C (VEGF-C) aggravated intestinal inflammation in mice with experimental acute colitis

    doi: 10.1590/1414-431X20154738

    Figure Lengend Snippet: Evaluation of disease activity index (DAI) in AD-VEGF-C-treated mice. Mean DAI scores (±SD) were higher in the AD-VEGF-C-treated mice ( A ) and in recombinant VEGF-C156S-treated mice ( B ), compared to DSS-treated mice. The difference in DAI scores between AD-VEGF-C-treated mice and DSS-treated mice became significant on the second day of observation and continued to differ thereafter. There was a significant difference in DAI scores between recombinant VEGF-C156S-treated mice and PBS-treated mice on days 3, 5, 6 and 7. *P

    Article Snippet: One limitation of the present study was that, due to the lack of a commercially available VEGF-C antibody, which can be used on murine specimens for confocal microscopy, we could not determine the identity of the cells that produced VEGF-C. Further studies are necessary to understand the role and functions of lymphangiogenesis in IBD.

    Techniques: Activity Assay, Mouse Assay, Recombinant

    G-Rh2 downregulated protein expression levels of VEGF-C, MMP9 and MMP2 in NSCLC cells. a-c A549 and co-cultured A549 with RAW264.7 derived M2 cells were treated with different concentrations of G-Rh2 for 24 h. Cell lysates were harvested. Protein expression levels of VEGF-C, MMP9 and MMP2 were examined by western blot. β-actin was used as a loading control. d - f )Quantification of VEGF-C, MMP9, and MMP-2 bands through quantification software. * P

    Journal: BMC Cancer

    Article Title: Modulation the crosstalk between tumor-associated macrophages and non-small cell lung cancer to inhibit tumor migration and invasion by ginsenoside Rh2

    doi: 10.1186/s12885-018-4299-4

    Figure Lengend Snippet: G-Rh2 downregulated protein expression levels of VEGF-C, MMP9 and MMP2 in NSCLC cells. a-c A549 and co-cultured A549 with RAW264.7 derived M2 cells were treated with different concentrations of G-Rh2 for 24 h. Cell lysates were harvested. Protein expression levels of VEGF-C, MMP9 and MMP2 were examined by western blot. β-actin was used as a loading control. d - f )Quantification of VEGF-C, MMP9, and MMP-2 bands through quantification software. * P

    Article Snippet: After deparaffinization, endogenous peroxidase activity was blocked by incubation with 3% peroxide-methanol solution at room temperature (RT) for 10 min, and then antigen retrieval was performed at 100 °C in an autoclave for 7 min. After washing with PBS, sections were incubated with primary antibodies against theCD206 monoclonal antibody (clone 10D6, Zhongshan Goldenbridge Biotechnology Co., LTD., Beijing, China) and VEGF-C (Santa Cruz Biotechnology, Santa Cruz, CA, USA)overnight at 4 °C.

    Techniques: Expressing, Cell Culture, Derivative Assay, Western Blot, Software

    G-Rh2 decreased VEGF-C and CD206 expression in vivo. a-d Female 5-week-old C57 mice ( n = 14) were subcutaneously injected with murine lung cancer cells. Then, they were randomly divided into two groups i.e. vehicle control and G-Rh2 treated groups. After 21 days treatment, tumors were taken out for immunohistochemical staining with VEGF-C and CD206. e Tumor size was measured daily. G-Rh2 administration reduced the tumor size. * P

    Journal: BMC Cancer

    Article Title: Modulation the crosstalk between tumor-associated macrophages and non-small cell lung cancer to inhibit tumor migration and invasion by ginsenoside Rh2

    doi: 10.1186/s12885-018-4299-4

    Figure Lengend Snippet: G-Rh2 decreased VEGF-C and CD206 expression in vivo. a-d Female 5-week-old C57 mice ( n = 14) were subcutaneously injected with murine lung cancer cells. Then, they were randomly divided into two groups i.e. vehicle control and G-Rh2 treated groups. After 21 days treatment, tumors were taken out for immunohistochemical staining with VEGF-C and CD206. e Tumor size was measured daily. G-Rh2 administration reduced the tumor size. * P

    Article Snippet: After deparaffinization, endogenous peroxidase activity was blocked by incubation with 3% peroxide-methanol solution at room temperature (RT) for 10 min, and then antigen retrieval was performed at 100 °C in an autoclave for 7 min. After washing with PBS, sections were incubated with primary antibodies against theCD206 monoclonal antibody (clone 10D6, Zhongshan Goldenbridge Biotechnology Co., LTD., Beijing, China) and VEGF-C (Santa Cruz Biotechnology, Santa Cruz, CA, USA)overnight at 4 °C.

    Techniques: Expressing, In Vivo, Mouse Assay, Injection, Immunohistochemistry, Staining

    Quantitative expression of VEGF-A, VEGF-C, VEGF-D according to the mode of tumour progression (A) For VEGF-A no significant differences in expression levels between the different groups could be detected. (B) VEGF-C with significantly higher expression in patients with ‘predominant retroperitoneal‘ metastases compared to ‘extensive intraperitoneal’ metastases (median 1.14 vs 0.42, p=0.001) and compared to patients with both types of metastases (median 1.14 vs 0.35, p=0.00002), as well as compared to patients with solely intraperitoneal metastases (median 1.14 vs 0.60, p=0.03). VEGF-C expression is significantly higher in the group of solely intraperitoneal metastases compared to both types of metastases (median 0.60 vs 0.35, p=0.007) (C) VEGF-D exhibits a trend of higher expression levels in patients with ’predominant retroperitoneal’ metastases compared to the ’extensive intraperitoneal’ group (mean 1.33 vs 0.96, p=0.09), although without statistical significance.

    Journal: Oncotarget

    Article Title: VEGF-C expression attributes the risk for lymphatic metastases to ovarian cancer patients

    doi: 10.18632/oncotarget.17978

    Figure Lengend Snippet: Quantitative expression of VEGF-A, VEGF-C, VEGF-D according to the mode of tumour progression (A) For VEGF-A no significant differences in expression levels between the different groups could be detected. (B) VEGF-C with significantly higher expression in patients with ‘predominant retroperitoneal‘ metastases compared to ‘extensive intraperitoneal’ metastases (median 1.14 vs 0.42, p=0.001) and compared to patients with both types of metastases (median 1.14 vs 0.35, p=0.00002), as well as compared to patients with solely intraperitoneal metastases (median 1.14 vs 0.60, p=0.03). VEGF-C expression is significantly higher in the group of solely intraperitoneal metastases compared to both types of metastases (median 0.60 vs 0.35, p=0.007) (C) VEGF-D exhibits a trend of higher expression levels in patients with ’predominant retroperitoneal’ metastases compared to the ’extensive intraperitoneal’ group (mean 1.33 vs 0.96, p=0.09), although without statistical significance.

    Article Snippet: The monoclonal antibody for VEGF-A (Abcam ab46154), 1:8000, Cambridge United Kingdom) was diluted in 5% nonfat dry milk in TBST, whereas the monoclonal antibodies for VEGF-C (sc374628, 1:200, Santa Cruz Biotechnology, Heidelberg, Germany) and VEGF-D (H144 sc13085, 1:1000, santa cruz) were diluted in 5% BSA in TBST.

    Techniques: Expressing

    Expression of VEGF-A, VEGF-C, VEGF-D according to the mode of tumour progression Representative expression of VEGF-A, VEGF-C, VEGF-D in the different types of tumour dissemination. Protein lysates from the breast cancer cell line MCF7 were used as positive controls for VEGF-A, –C and –D. Equal amounts of protein lysate (20 μg) were loaded per well.

    Journal: Oncotarget

    Article Title: VEGF-C expression attributes the risk for lymphatic metastases to ovarian cancer patients

    doi: 10.18632/oncotarget.17978

    Figure Lengend Snippet: Expression of VEGF-A, VEGF-C, VEGF-D according to the mode of tumour progression Representative expression of VEGF-A, VEGF-C, VEGF-D in the different types of tumour dissemination. Protein lysates from the breast cancer cell line MCF7 were used as positive controls for VEGF-A, –C and –D. Equal amounts of protein lysate (20 μg) were loaded per well.

    Article Snippet: The monoclonal antibody for VEGF-A (Abcam ab46154), 1:8000, Cambridge United Kingdom) was diluted in 5% nonfat dry milk in TBST, whereas the monoclonal antibodies for VEGF-C (sc374628, 1:200, Santa Cruz Biotechnology, Heidelberg, Germany) and VEGF-D (H144 sc13085, 1:1000, santa cruz) were diluted in 5% BSA in TBST.

    Techniques: Expressing

    Progression-free and overall survival according to the expression level of VEGF-C By dividing the overall patient cohort into two groups according to the median VEGF-C expression by Western Blot, two groups with low (n=50) and high (n=50) VEGF-C expression were generated. (A) Progression-free survival shows no prognostic differences in relation to VEGF-C expression (median 23 vs 23 months; HR1.44, 95%-CI 0.89-2.31, p=0.13; Log Rank p=0.13). (B) Patients with high VEGF-C expression have a significantly shorter overall survival compared to patients with low VEGF-C expression levels with a median of 41 versus 56 months (HR 2.02, 95%-CI 1.12-3.63, p=0.019; Log Rank p=0.016).

    Journal: Oncotarget

    Article Title: VEGF-C expression attributes the risk for lymphatic metastases to ovarian cancer patients

    doi: 10.18632/oncotarget.17978

    Figure Lengend Snippet: Progression-free and overall survival according to the expression level of VEGF-C By dividing the overall patient cohort into two groups according to the median VEGF-C expression by Western Blot, two groups with low (n=50) and high (n=50) VEGF-C expression were generated. (A) Progression-free survival shows no prognostic differences in relation to VEGF-C expression (median 23 vs 23 months; HR1.44, 95%-CI 0.89-2.31, p=0.13; Log Rank p=0.13). (B) Patients with high VEGF-C expression have a significantly shorter overall survival compared to patients with low VEGF-C expression levels with a median of 41 versus 56 months (HR 2.02, 95%-CI 1.12-3.63, p=0.019; Log Rank p=0.016).

    Article Snippet: The monoclonal antibody for VEGF-A (Abcam ab46154), 1:8000, Cambridge United Kingdom) was diluted in 5% nonfat dry milk in TBST, whereas the monoclonal antibodies for VEGF-C (sc374628, 1:200, Santa Cruz Biotechnology, Heidelberg, Germany) and VEGF-D (H144 sc13085, 1:1000, santa cruz) were diluted in 5% BSA in TBST.

    Techniques: Expressing, Western Blot, Generated

    Expression patterns determining different types of metastases in EOC Tumour cells of patients with ‘predominant retroperitoneal‘ metastases are characterised by a mesenchymal tumour type with low E-Cadherin and high VEGF-C and –D expression, which probably leads to local intrapelvic tumour growth and lymphatic metastases instead of disseminating diffusely intraabdominal. The tumour type of patients with solely intraperitoneal metastases is otherwise characterised by an epithelial phenotype with high E-Cadherin and low VEGF-C and -D expression, which potentially leads to ‘extensive intraperitoneal’ tumour dissemination (A: ascites, LV: lymph-vessel, TC: tumour cell).

    Journal: Oncotarget

    Article Title: VEGF-C expression attributes the risk for lymphatic metastases to ovarian cancer patients

    doi: 10.18632/oncotarget.17978

    Figure Lengend Snippet: Expression patterns determining different types of metastases in EOC Tumour cells of patients with ‘predominant retroperitoneal‘ metastases are characterised by a mesenchymal tumour type with low E-Cadherin and high VEGF-C and –D expression, which probably leads to local intrapelvic tumour growth and lymphatic metastases instead of disseminating diffusely intraabdominal. The tumour type of patients with solely intraperitoneal metastases is otherwise characterised by an epithelial phenotype with high E-Cadherin and low VEGF-C and -D expression, which potentially leads to ‘extensive intraperitoneal’ tumour dissemination (A: ascites, LV: lymph-vessel, TC: tumour cell).

    Article Snippet: The monoclonal antibody for VEGF-A (Abcam ab46154), 1:8000, Cambridge United Kingdom) was diluted in 5% nonfat dry milk in TBST, whereas the monoclonal antibodies for VEGF-C (sc374628, 1:200, Santa Cruz Biotechnology, Heidelberg, Germany) and VEGF-D (H144 sc13085, 1:1000, santa cruz) were diluted in 5% BSA in TBST.

    Techniques: Expressing