vascular endothelial growth factor vegf a  (Thermo Fisher)


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    • 99
    Name:
    Vascular Endothelial Growth Factor VEGF Ab 7
    Description:

    Catalog Number:
    ms-1467-pcl
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    Structured Review

    Thermo Fisher vascular endothelial growth factor vegf a
    Impaired chronic allergen-induced airway remodeling and collagen deposition in sPLA 2 -V −/− mice. Lung tissue was obtained on d 76 from sPLA 2 -V +/+ and sPLA 2 -V −/− mice treated with either saline or OVA. A. Sections underwent Masson's trichrome staining. Scale bar, 50 μm. Micrographs are representative from 4–5 mice per group. B. Airway subepithelial fibrosis (0–4+ scale) was determined by morphometry, and lung collagen deposition (μg/lung) was determined by Sircol™ assay ( n = 4–5, each group). C. Collagen (COL1α2 and COL3α1) and <t>VEGF</t> <t>(VEGF-A,</t> VEGF-A2, VEGF-B, and VEGF-C) gene expression in OVA-treated sPLA 2 -V +/+ (black bars) and sPLA 2 -V −/− (blue bars) mice was determined by qPCR; nd = not detected in samples from the sPLA 2 -V −/− mice. * P

    https://www.bioz.com/result/vascular endothelial growth factor vegf a/product/Thermo Fisher
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    Images

    1) Product Images from "Key Role of Group V Secreted Phospholipase A2 in Th2 Cytokine and Dendritic Cell-Driven Airway Hyperresponsiveness and Remodeling"

    Article Title: Key Role of Group V Secreted Phospholipase A2 in Th2 Cytokine and Dendritic Cell-Driven Airway Hyperresponsiveness and Remodeling

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0056172

    Impaired chronic allergen-induced airway remodeling and collagen deposition in sPLA 2 -V −/− mice. Lung tissue was obtained on d 76 from sPLA 2 -V +/+ and sPLA 2 -V −/− mice treated with either saline or OVA. A. Sections underwent Masson's trichrome staining. Scale bar, 50 μm. Micrographs are representative from 4–5 mice per group. B. Airway subepithelial fibrosis (0–4+ scale) was determined by morphometry, and lung collagen deposition (μg/lung) was determined by Sircol™ assay ( n = 4–5, each group). C. Collagen (COL1α2 and COL3α1) and VEGF (VEGF-A, VEGF-A2, VEGF-B, and VEGF-C) gene expression in OVA-treated sPLA 2 -V +/+ (black bars) and sPLA 2 -V −/− (blue bars) mice was determined by qPCR; nd = not detected in samples from the sPLA 2 -V −/− mice. * P
    Figure Legend Snippet: Impaired chronic allergen-induced airway remodeling and collagen deposition in sPLA 2 -V −/− mice. Lung tissue was obtained on d 76 from sPLA 2 -V +/+ and sPLA 2 -V −/− mice treated with either saline or OVA. A. Sections underwent Masson's trichrome staining. Scale bar, 50 μm. Micrographs are representative from 4–5 mice per group. B. Airway subepithelial fibrosis (0–4+ scale) was determined by morphometry, and lung collagen deposition (μg/lung) was determined by Sircol™ assay ( n = 4–5, each group). C. Collagen (COL1α2 and COL3α1) and VEGF (VEGF-A, VEGF-A2, VEGF-B, and VEGF-C) gene expression in OVA-treated sPLA 2 -V +/+ (black bars) and sPLA 2 -V −/− (blue bars) mice was determined by qPCR; nd = not detected in samples from the sPLA 2 -V −/− mice. * P

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

    2) Product Images from "Three-dimensional spheroid cell culture of umbilical cord tissue-derived mesenchymal stromal cells leads to enhanced paracrine induction of wound healing"

    Article Title: Three-dimensional spheroid cell culture of umbilical cord tissue-derived mesenchymal stromal cells leads to enhanced paracrine induction of wound healing

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-015-0082-5

    The secretion of growth factors involved in the wound-healing process was increased in UCX® spheroids. Hepatocyte growth factor (HGF), transforming growth factor (TGF)-β1, granulocyte-colony stimulating factor (G-CSF), vascular endothelial growth factor (VEGF)-A, fibroblast growth factor (FGF)-2 and interleukin (IL)-6 were quantified in two-dimensional conditioned medium (CM2D) and three-dimensional conditioned medium (CM3D) using a FlowCytomix™ kit. Keratinocyte growth factor (KGF) was quantified in CM2D and CM3D by enzyme-linked immunosorbent assay. Results are expressed as fold-increase of CM3D relative to CM2D. Data are shown in mean ± standard deviation; n = 3.
    Figure Legend Snippet: The secretion of growth factors involved in the wound-healing process was increased in UCX® spheroids. Hepatocyte growth factor (HGF), transforming growth factor (TGF)-β1, granulocyte-colony stimulating factor (G-CSF), vascular endothelial growth factor (VEGF)-A, fibroblast growth factor (FGF)-2 and interleukin (IL)-6 were quantified in two-dimensional conditioned medium (CM2D) and three-dimensional conditioned medium (CM3D) using a FlowCytomix™ kit. Keratinocyte growth factor (KGF) was quantified in CM2D and CM3D by enzyme-linked immunosorbent assay. Results are expressed as fold-increase of CM3D relative to CM2D. Data are shown in mean ± standard deviation; n = 3.

    Techniques Used: Enzyme-linked Immunosorbent Assay, Standard Deviation

    Related Articles

    Enzyme-linked Immunosorbent Assay:

    Article Title: Effects of Peroxisome Proliferator-Activated Receptor-δ Agonist on Cardiac Healing after Myocardial Infarction
    Article Snippet: .. Measurement of serum platelet-derived growth factor B (PDGF-b), matrix metallopeptidase 9 (MMP-9), basic fibroblast growth factor (bFGF), stromal-derived factor-1 alpha (SDF-1α), and vascular endothelial growth factor (VEGF) Serum levels of PDGF-b, MMP-9, bFGF, SDF-1α and VEGF were quantified using specific enzyme-linked immunosorbent assay kits (ELISA) according to the manufacturer’s instructions (Biosource International, Nivelles, Belgium). ..

    Article Title: Macrophage migratory inhibitory factor promotes bladder cancer progression via increasing proliferation and angiogenesis
    Article Snippet: Bands were scanned for images and densitometry for phosphorylated/total ERK and phosphorylated/total AKT was performed using NIH Image J software (National Institutes of Health, Bethesda, MD). .. Medium was removed from cultured HTB-5 cells, treated ± CPSI rhMIF (100ng/ml) ± CPS1-1306 (50nM), and vascular endothelial growth factor (VEGF) secretion was measured using HUMAN VEGF ELISA KIT (Invitrogen, Life Technologies) following manufacturer’s guidelines. .. C57Bl/6 mice were purchased from Charles River (Wilmington, MA) and maintained at the University of Connecticut Health Center for Laboratory Animal Care under National Institutes of Health guidelines.

    Cell Culture:

    Article Title: Macrophage migratory inhibitory factor promotes bladder cancer progression via increasing proliferation and angiogenesis
    Article Snippet: Bands were scanned for images and densitometry for phosphorylated/total ERK and phosphorylated/total AKT was performed using NIH Image J software (National Institutes of Health, Bethesda, MD). .. Medium was removed from cultured HTB-5 cells, treated ± CPSI rhMIF (100ng/ml) ± CPS1-1306 (50nM), and vascular endothelial growth factor (VEGF) secretion was measured using HUMAN VEGF ELISA KIT (Invitrogen, Life Technologies) following manufacturer’s guidelines. .. C57Bl/6 mice were purchased from Charles River (Wilmington, MA) and maintained at the University of Connecticut Health Center for Laboratory Animal Care under National Institutes of Health guidelines.

    Real-time Polymerase Chain Reaction:

    Article Title: Tongxinluo Improves Apolipoprotein E-Deficient Mouse Heart Function
    Article Snippet: .. Real-time quantitative polymerase chain reaction and Western blotting analysis of vascular endothelial growth factor expression To detect the vascular endothelial growth factor (VEGF ) mRNA level, total RNA from heart tissues was extracted with Trizol (Invitrogen, America) and purified (Qiagen, Germany). .. The mRNAs were reverse transcribed into cDNAs with a SuperScript cDNA synthesis kit (Invitrogen, America).

    Article Title: Adipose-Derived Mesenchymal Stem Cells Migrate and Rescue RPE in the Setting of Oxidative Stress
    Article Snippet: Total RNA concentration was determined by NanoDrop™ 1000 Spectrophotometer (Thermo Scientific) and was reverse transcribed using Verso cDNA synthesis kit (Thermo Scientific). .. The mRNA expression levels of the growth factors, hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), interleukin-1 β (IL-1 β ), stromal-derived factor-1 (SDF-1), the chemokine receptor CXCR4, and normalizing housekeeping genes GUSB and RLP27 (see for sequence information) were measured by real-time reverse transcription polymerase chain reaction (RT-PCR) (StepOnePlus, Applied Biosystems) using SYBR® Green qPCR Mastermix (Qiagen). .. The cycling RT-PCR conditions were as follows: 10 min at 95°C, 40 cycles for 10 s at 95°C, 15 s at 60°C, followed by gradient stage from 60 to 95°C to obtain a melting curve.

    Western Blot:

    Article Title: Tongxinluo Improves Apolipoprotein E-Deficient Mouse Heart Function
    Article Snippet: .. Real-time quantitative polymerase chain reaction and Western blotting analysis of vascular endothelial growth factor expression To detect the vascular endothelial growth factor (VEGF ) mRNA level, total RNA from heart tissues was extracted with Trizol (Invitrogen, America) and purified (Qiagen, Germany). .. The mRNAs were reverse transcribed into cDNAs with a SuperScript cDNA synthesis kit (Invitrogen, America).

    Expressing:

    Article Title: Tongxinluo Improves Apolipoprotein E-Deficient Mouse Heart Function
    Article Snippet: .. Real-time quantitative polymerase chain reaction and Western blotting analysis of vascular endothelial growth factor expression To detect the vascular endothelial growth factor (VEGF ) mRNA level, total RNA from heart tissues was extracted with Trizol (Invitrogen, America) and purified (Qiagen, Germany). .. The mRNAs were reverse transcribed into cDNAs with a SuperScript cDNA synthesis kit (Invitrogen, America).

    Article Title: Prognostic significance of Cytokeratin 20-positive lymph node vascular endothelial growth factor A mRNA and chromodomain helicase DNA binding protein 4 in pN0 colorectal cancer patients
    Article Snippet: .. Validation of vascular endothelial growth factor A (VEGF-A) mRNA and transcription factor 20 (TCF20) mRNA in the second cohort of PELS TaqMan gene expression assays of specific primers and MGB probes were purchased from Applied Biosystems for the detection of VEGF-A mRNA (Hs00900054_m1) and TCF20 mRNA (Hs00390028_m1). .. QRT-PCR was performed in a reaction volume of 50 μL using Taqman Universal PCR Master Mix (Cat. no. 4304437; Applied Biosystems), and 10 μL of cDNA was used for each reaction.

    Article Title: Inhibition of androgen-responsive LNCaP prostate cancer cell tumor xenograft growth by dietary phenethyl isothiocyanate correlates with decreased angiogenesis and inhibition of cell attachment
    Article Snippet: Gene expression was quantified by the TaqMan real-time RT-PCR method as described previously ( ). .. TaqMan gene expression assay primers and probes for human prostate specific antigen (PSA) and glyceraldehydes-3-phosphate dehydrogenase (G3PDH), proliferating cell nuclear antigen (PCNA), Ki-67, integrin β1 (ITGB1), integrin α2 (ITGA2), integrin α5 (ITGA5), integrin α6 (ITGA6), mouse and human vascular endothelial growth factor (VEGF) A, mouse PECAM-1 and mouse and human TATA binding protein were purchased from Applied Biosystems (Foster City, CA). .. G3PDH was used as a housekeeping gene for calculation of relative expression levels in vitro and mouse or human TATA binding protein was used for tumor samples.

    Article Title: Adipose-Derived Mesenchymal Stem Cells Migrate and Rescue RPE in the Setting of Oxidative Stress
    Article Snippet: Total RNA concentration was determined by NanoDrop™ 1000 Spectrophotometer (Thermo Scientific) and was reverse transcribed using Verso cDNA synthesis kit (Thermo Scientific). .. The mRNA expression levels of the growth factors, hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), interleukin-1 β (IL-1 β ), stromal-derived factor-1 (SDF-1), the chemokine receptor CXCR4, and normalizing housekeeping genes GUSB and RLP27 (see for sequence information) were measured by real-time reverse transcription polymerase chain reaction (RT-PCR) (StepOnePlus, Applied Biosystems) using SYBR® Green qPCR Mastermix (Qiagen). .. The cycling RT-PCR conditions were as follows: 10 min at 95°C, 40 cycles for 10 s at 95°C, 15 s at 60°C, followed by gradient stage from 60 to 95°C to obtain a melting curve.

    Purification:

    Article Title: Tongxinluo Improves Apolipoprotein E-Deficient Mouse Heart Function
    Article Snippet: .. Real-time quantitative polymerase chain reaction and Western blotting analysis of vascular endothelial growth factor expression To detect the vascular endothelial growth factor (VEGF ) mRNA level, total RNA from heart tissues was extracted with Trizol (Invitrogen, America) and purified (Qiagen, Germany). .. The mRNAs were reverse transcribed into cDNAs with a SuperScript cDNA synthesis kit (Invitrogen, America).

    Multiplex Assay:

    Article Title: Correlation between thrombocytopenia and host response in severe fever with thrombocytopenia syndrome
    Article Snippet: Briefly, the cytokine/chemokines were evaluated from the acute serum samples by the Bio-plex Pro Human 27-plex cytokine panel (Bio-Rad) following manufacturer’s instruction. .. Serum levels of vascular endothelial growth factor A (VEGF-A), P-selectin, E-selectin, platelet endothelial cell adhesion molecular (PECAM-1), CD40 ligand (CD40L), tissue plasminogen activator (tPA), plasminogen activator inhibitor 1 (PAI-1), serum amyloid antigen 1 (SAA-1), vascular cell adhesion molecular 1 (VCAM-1) and intercellular adhesion molecular 1 (ICAM-1) were determined by the ProcartaPlex multiplex immunoassays panels (Affymetrix, USA) according to the manufacturer instructions. ..

    Binding Assay:

    Article Title: Inhibition of androgen-responsive LNCaP prostate cancer cell tumor xenograft growth by dietary phenethyl isothiocyanate correlates with decreased angiogenesis and inhibition of cell attachment
    Article Snippet: Gene expression was quantified by the TaqMan real-time RT-PCR method as described previously ( ). .. TaqMan gene expression assay primers and probes for human prostate specific antigen (PSA) and glyceraldehydes-3-phosphate dehydrogenase (G3PDH), proliferating cell nuclear antigen (PCNA), Ki-67, integrin β1 (ITGB1), integrin α2 (ITGA2), integrin α5 (ITGA5), integrin α6 (ITGA6), mouse and human vascular endothelial growth factor (VEGF) A, mouse PECAM-1 and mouse and human TATA binding protein were purchased from Applied Biosystems (Foster City, CA). .. G3PDH was used as a housekeeping gene for calculation of relative expression levels in vitro and mouse or human TATA binding protein was used for tumor samples.

    Sequencing:

    Article Title: Adipose-Derived Mesenchymal Stem Cells Migrate and Rescue RPE in the Setting of Oxidative Stress
    Article Snippet: Total RNA concentration was determined by NanoDrop™ 1000 Spectrophotometer (Thermo Scientific) and was reverse transcribed using Verso cDNA synthesis kit (Thermo Scientific). .. The mRNA expression levels of the growth factors, hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), interleukin-1 β (IL-1 β ), stromal-derived factor-1 (SDF-1), the chemokine receptor CXCR4, and normalizing housekeeping genes GUSB and RLP27 (see for sequence information) were measured by real-time reverse transcription polymerase chain reaction (RT-PCR) (StepOnePlus, Applied Biosystems) using SYBR® Green qPCR Mastermix (Qiagen). .. The cycling RT-PCR conditions were as follows: 10 min at 95°C, 40 cycles for 10 s at 95°C, 15 s at 60°C, followed by gradient stage from 60 to 95°C to obtain a melting curve.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Adipose-Derived Mesenchymal Stem Cells Migrate and Rescue RPE in the Setting of Oxidative Stress
    Article Snippet: Total RNA concentration was determined by NanoDrop™ 1000 Spectrophotometer (Thermo Scientific) and was reverse transcribed using Verso cDNA synthesis kit (Thermo Scientific). .. The mRNA expression levels of the growth factors, hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), interleukin-1 β (IL-1 β ), stromal-derived factor-1 (SDF-1), the chemokine receptor CXCR4, and normalizing housekeeping genes GUSB and RLP27 (see for sequence information) were measured by real-time reverse transcription polymerase chain reaction (RT-PCR) (StepOnePlus, Applied Biosystems) using SYBR® Green qPCR Mastermix (Qiagen). .. The cycling RT-PCR conditions were as follows: 10 min at 95°C, 40 cycles for 10 s at 95°C, 15 s at 60°C, followed by gradient stage from 60 to 95°C to obtain a melting curve.

    SYBR Green Assay:

    Article Title: Adipose-Derived Mesenchymal Stem Cells Migrate and Rescue RPE in the Setting of Oxidative Stress
    Article Snippet: Total RNA concentration was determined by NanoDrop™ 1000 Spectrophotometer (Thermo Scientific) and was reverse transcribed using Verso cDNA synthesis kit (Thermo Scientific). .. The mRNA expression levels of the growth factors, hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), interleukin-1 β (IL-1 β ), stromal-derived factor-1 (SDF-1), the chemokine receptor CXCR4, and normalizing housekeeping genes GUSB and RLP27 (see for sequence information) were measured by real-time reverse transcription polymerase chain reaction (RT-PCR) (StepOnePlus, Applied Biosystems) using SYBR® Green qPCR Mastermix (Qiagen). .. The cycling RT-PCR conditions were as follows: 10 min at 95°C, 40 cycles for 10 s at 95°C, 15 s at 60°C, followed by gradient stage from 60 to 95°C to obtain a melting curve.

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    • gene exp flt4 mm01292618 m1  (Thermo Fisher)
    92
    Thermo Fisher gene exp flt4 mm01292618 m1
    NMDA-Triggered Excitotoxicity Reduces VEGFD Expression in RGCs (A) qRT-PCR analysis of vegfd , vegfc , vegfa , <t>flt4</t> , flt1 , and kdr expression in retinal homogenates after intravitreal injection of NMDA. Unpaired t test. n = 6. vegfd , p = 0.0085; vegfc , p = 0.2052; vegfa , p = 0.8524; flt4 , p = 0.9414; flt1 , p = 0.6074; kdr , p = 0.2016. (B) Representative images of human retinal whole mounts. Retinas were immunolabeled with antibodies against Brn3a (red) and VEGFD (green). Scale bars, 10 μm. Negative controls of human retinas were immunolabeled only with secondary antibodies. (C) Representative bands of VEGFD, VEGFA, and β-actin cDNA products following qRT-PCR analysis of human retinal extracts obtained from two donors (subject 1, subject 2). (D) Quantification of VEGFD protein levels in cells in the ganglion cell layer of retinas of mice injected as indicated. Unpaired t test. n = 3. PBS versus NMDA, p = 0.0192. (E) Representative images of mouse sagittal retina sections at d7 after intravitreal injections of NMDA or PBS. Retinas were immunolabeled for VEGFD (green). Nuclei were labeled with Hoechst (blue). GCL, ganglion cell layer. Scale bar, 20 μm. (F) qRT-PCR analysis of vegfd , vegfc , vegfa , and flt4 expression in brain ECs (b.END3) 24 h after 20 μM NMDA treatment. Unpaired t test. n = 3. vegfd , p = 0.1744; vegfc , p = 0.7598; vegfa , p = 0.8998; flt4 , p = 0.4457. (G) qRT-PCR analysis of vegfd , vegfc , vegfa , and flt4 expression in primary astrocytes 24 h after 20 μM NMDA treatment. Unpaired t test. n = 3. vegfd , p = 0.8716; vegfc , p = 0.4918; vegfa , p = 0.9902; flt4 , p = 0.9331. Bars represent mean ± SEM. Dots represent single values. **p
    Gene Exp Flt4 Mm01292618 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gene exp flt4 mm01292618 m1/product/Thermo Fisher
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    gene exp flt4 mm01292618 m1 - by Bioz Stars, 2021-03
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    gene exp vegfa mm01281449 m1  (Thermo Fisher)
    99
    Thermo Fisher gene exp vegfa mm01281449 m1
    NMDA-Triggered Excitotoxicity Reduces VEGFD Expression in RGCs (A) qRT-PCR analysis of vegfd , vegfc , <t>vegfa</t> , flt4 , flt1 , and kdr expression in retinal homogenates after intravitreal injection of NMDA. Unpaired t test. n = 6. vegfd , p = 0.0085; vegfc , p = 0.2052; vegfa , p = 0.8524; flt4 , p = 0.9414; flt1 , p = 0.6074; kdr , p = 0.2016. (B) Representative images of human retinal whole mounts. Retinas were immunolabeled with antibodies against Brn3a (red) and VEGFD (green). Scale bars, 10 μm. Negative controls of human retinas were immunolabeled only with secondary antibodies. (C) Representative bands of VEGFD, VEGFA, and β-actin cDNA products following qRT-PCR analysis of human retinal extracts obtained from two donors (subject 1, subject 2). (D) Quantification of VEGFD protein levels in cells in the ganglion cell layer of retinas of mice injected as indicated. Unpaired t test. n = 3. PBS versus NMDA, p = 0.0192. (E) Representative images of mouse sagittal retina sections at d7 after intravitreal injections of NMDA or PBS. Retinas were immunolabeled for VEGFD (green). Nuclei were labeled with Hoechst (blue). GCL, ganglion cell layer. Scale bar, 20 μm. (F) qRT-PCR analysis of vegfd , vegfc , vegfa , and flt4 expression in brain ECs (b.END3) 24 h after 20 μM NMDA treatment. Unpaired t test. n = 3. vegfd , p = 0.1744; vegfc , p = 0.7598; vegfa , p = 0.8998; flt4 , p = 0.4457. (G) qRT-PCR analysis of vegfd , vegfc , vegfa , and flt4 expression in primary astrocytes 24 h after 20 μM NMDA treatment. Unpaired t test. n = 3. vegfd , p = 0.8716; vegfc , p = 0.4918; vegfa , p = 0.9902; flt4 , p = 0.9331. Bars represent mean ± SEM. Dots represent single values. **p
    Gene Exp Vegfa Mm01281449 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gene exp vegfa mm01281449 m1/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
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    gene exp vegfa mm00437306 m1  (Thermo Fisher)
    99
    Thermo Fisher gene exp vegfa mm00437306 m1
    A. Gating strategy for analyzing Sema3A expression among leukocytes, epithelial and endothelial cells in lung and dLN. B. Quantification of Sema3A positive cells in different cell populations in uninfected (n=4), and infected mice, at 3 days DPI (n=4) and 10 DPI (n=4) in lung (left panel) or dLN (right panel). Nrp1 Flox/Flox mice used. Data combined from two independent experiments. ns = not significant by two-way ANOVA. C. Quantification of Pkd1, Bnip3, <t>Vegfa,</t> Pdl1 and Sema3A mRNA level following 24 hour culture in 1% O2 chamber. D. Gating strategy (left) and representative histograms (right) analyzing Sema3A positive cell populations in B16.F10 tumors 11 days post-injection. Olfactory lobe is used as a positive control for Sema3A expression. E. Quantification of Sema3A positive cells in same experiment as in (D) in tumor, dLN and ndLN. F. Genomic organization of murine Sema3a gene, indicating where CRISPR guide RNA targets with red arrow (upper figure). MiSEQ sequence results for chosen Sema3A KO clone showing 4 base deletion in two alleles (46% of all reads), a frameshift in one allele (45% of all reads) and WT reads in 2% of all reads. G. RT-qPCR analysis show down- and up-regulation of Sema3A in Sema3A KO and OE cell lines, respectively. Normalized to Hprt. Experiment performed once. H. Intracellular staining shows no detectable expression of Sema3A in Sema3A KO cells, and expression in Sema3A OE cells, as expected. Experiment performed once, at low seeding density. I. Growth of WT, Sema3A KO and Sema3A OE B16.F10.Ova cell lines in normal, IFNγ or TNFα-rich media. Experiment performed once. Data indicate mean ± SD. ns = not significant by two-way ANOVA. Abbreviations: BEC, blood endothelial cells. dLN, draining lymph node. DPI, days post-infection. FRC, fibroblastic reticular cells. KO, knockout. LEC, lymphatic endothelial cells. ndLN, non-draining lymph node. OE, overexpressing. TME, tumor microenvironment. WT, wild-type.
    Gene Exp Vegfa Mm00437306 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    NMDA-Triggered Excitotoxicity Reduces VEGFD Expression in RGCs (A) qRT-PCR analysis of vegfd , vegfc , vegfa , flt4 , flt1 , and kdr expression in retinal homogenates after intravitreal injection of NMDA. Unpaired t test. n = 6. vegfd , p = 0.0085; vegfc , p = 0.2052; vegfa , p = 0.8524; flt4 , p = 0.9414; flt1 , p = 0.6074; kdr , p = 0.2016. (B) Representative images of human retinal whole mounts. Retinas were immunolabeled with antibodies against Brn3a (red) and VEGFD (green). Scale bars, 10 μm. Negative controls of human retinas were immunolabeled only with secondary antibodies. (C) Representative bands of VEGFD, VEGFA, and β-actin cDNA products following qRT-PCR analysis of human retinal extracts obtained from two donors (subject 1, subject 2). (D) Quantification of VEGFD protein levels in cells in the ganglion cell layer of retinas of mice injected as indicated. Unpaired t test. n = 3. PBS versus NMDA, p = 0.0192. (E) Representative images of mouse sagittal retina sections at d7 after intravitreal injections of NMDA or PBS. Retinas were immunolabeled for VEGFD (green). Nuclei were labeled with Hoechst (blue). GCL, ganglion cell layer. Scale bar, 20 μm. (F) qRT-PCR analysis of vegfd , vegfc , vegfa , and flt4 expression in brain ECs (b.END3) 24 h after 20 μM NMDA treatment. Unpaired t test. n = 3. vegfd , p = 0.1744; vegfc , p = 0.7598; vegfa , p = 0.8998; flt4 , p = 0.4457. (G) qRT-PCR analysis of vegfd , vegfc , vegfa , and flt4 expression in primary astrocytes 24 h after 20 μM NMDA treatment. Unpaired t test. n = 3. vegfd , p = 0.8716; vegfc , p = 0.4918; vegfa , p = 0.9902; flt4 , p = 0.9331. Bars represent mean ± SEM. Dots represent single values. **p

    Journal: Molecular Therapy. Methods & Clinical Development

    Article Title: VEGFD Protects Retinal Ganglion Cells and, consequently, Capillaries against Excitotoxic Injury

    doi: 10.1016/j.omtm.2019.12.009

    Figure Lengend Snippet: NMDA-Triggered Excitotoxicity Reduces VEGFD Expression in RGCs (A) qRT-PCR analysis of vegfd , vegfc , vegfa , flt4 , flt1 , and kdr expression in retinal homogenates after intravitreal injection of NMDA. Unpaired t test. n = 6. vegfd , p = 0.0085; vegfc , p = 0.2052; vegfa , p = 0.8524; flt4 , p = 0.9414; flt1 , p = 0.6074; kdr , p = 0.2016. (B) Representative images of human retinal whole mounts. Retinas were immunolabeled with antibodies against Brn3a (red) and VEGFD (green). Scale bars, 10 μm. Negative controls of human retinas were immunolabeled only with secondary antibodies. (C) Representative bands of VEGFD, VEGFA, and β-actin cDNA products following qRT-PCR analysis of human retinal extracts obtained from two donors (subject 1, subject 2). (D) Quantification of VEGFD protein levels in cells in the ganglion cell layer of retinas of mice injected as indicated. Unpaired t test. n = 3. PBS versus NMDA, p = 0.0192. (E) Representative images of mouse sagittal retina sections at d7 after intravitreal injections of NMDA or PBS. Retinas were immunolabeled for VEGFD (green). Nuclei were labeled with Hoechst (blue). GCL, ganglion cell layer. Scale bar, 20 μm. (F) qRT-PCR analysis of vegfd , vegfc , vegfa , and flt4 expression in brain ECs (b.END3) 24 h after 20 μM NMDA treatment. Unpaired t test. n = 3. vegfd , p = 0.1744; vegfc , p = 0.7598; vegfa , p = 0.8998; flt4 , p = 0.4457. (G) qRT-PCR analysis of vegfd , vegfc , vegfa , and flt4 expression in primary astrocytes 24 h after 20 μM NMDA treatment. Unpaired t test. n = 3. vegfd , p = 0.8716; vegfc , p = 0.4918; vegfa , p = 0.9902; flt4 , p = 0.9331. Bars represent mean ± SEM. Dots represent single values. **p

    Article Snippet: The following TaqMan gene expression assays were used in this study: Thy1 (Mm01174153_m1), nefl (Mm01315666_m1), ocld (Mm01282968_m1), cld5 (Mm00727012_m1), tjp1 (Mm00493699_s1), vegfd (Mm00438963_m1), vegfc (Mm01202432_m1), vegf (Mm01281449_m1), flt4 (Mm01292618_m1), flt1 (Mm00438980_m1), and kdr (Mm00440099_m1).

    Techniques: Expressing, Quantitative RT-PCR, Injection, Immunolabeling, Mouse Assay, Labeling

    NMDA-Triggered Excitotoxicity Reduces VEGFD Expression in RGCs (A) qRT-PCR analysis of vegfd , vegfc , vegfa , flt4 , flt1 , and kdr expression in retinal homogenates after intravitreal injection of NMDA. Unpaired t test. n = 6. vegfd , p = 0.0085; vegfc , p = 0.2052; vegfa , p = 0.8524; flt4 , p = 0.9414; flt1 , p = 0.6074; kdr , p = 0.2016. (B) Representative images of human retinal whole mounts. Retinas were immunolabeled with antibodies against Brn3a (red) and VEGFD (green). Scale bars, 10 μm. Negative controls of human retinas were immunolabeled only with secondary antibodies. (C) Representative bands of VEGFD, VEGFA, and β-actin cDNA products following qRT-PCR analysis of human retinal extracts obtained from two donors (subject 1, subject 2). (D) Quantification of VEGFD protein levels in cells in the ganglion cell layer of retinas of mice injected as indicated. Unpaired t test. n = 3. PBS versus NMDA, p = 0.0192. (E) Representative images of mouse sagittal retina sections at d7 after intravitreal injections of NMDA or PBS. Retinas were immunolabeled for VEGFD (green). Nuclei were labeled with Hoechst (blue). GCL, ganglion cell layer. Scale bar, 20 μm. (F) qRT-PCR analysis of vegfd , vegfc , vegfa , and flt4 expression in brain ECs (b.END3) 24 h after 20 μM NMDA treatment. Unpaired t test. n = 3. vegfd , p = 0.1744; vegfc , p = 0.7598; vegfa , p = 0.8998; flt4 , p = 0.4457. (G) qRT-PCR analysis of vegfd , vegfc , vegfa , and flt4 expression in primary astrocytes 24 h after 20 μM NMDA treatment. Unpaired t test. n = 3. vegfd , p = 0.8716; vegfc , p = 0.4918; vegfa , p = 0.9902; flt4 , p = 0.9331. Bars represent mean ± SEM. Dots represent single values. **p

    Journal: Molecular Therapy. Methods & Clinical Development

    Article Title: VEGFD Protects Retinal Ganglion Cells and, consequently, Capillaries against Excitotoxic Injury

    doi: 10.1016/j.omtm.2019.12.009

    Figure Lengend Snippet: NMDA-Triggered Excitotoxicity Reduces VEGFD Expression in RGCs (A) qRT-PCR analysis of vegfd , vegfc , vegfa , flt4 , flt1 , and kdr expression in retinal homogenates after intravitreal injection of NMDA. Unpaired t test. n = 6. vegfd , p = 0.0085; vegfc , p = 0.2052; vegfa , p = 0.8524; flt4 , p = 0.9414; flt1 , p = 0.6074; kdr , p = 0.2016. (B) Representative images of human retinal whole mounts. Retinas were immunolabeled with antibodies against Brn3a (red) and VEGFD (green). Scale bars, 10 μm. Negative controls of human retinas were immunolabeled only with secondary antibodies. (C) Representative bands of VEGFD, VEGFA, and β-actin cDNA products following qRT-PCR analysis of human retinal extracts obtained from two donors (subject 1, subject 2). (D) Quantification of VEGFD protein levels in cells in the ganglion cell layer of retinas of mice injected as indicated. Unpaired t test. n = 3. PBS versus NMDA, p = 0.0192. (E) Representative images of mouse sagittal retina sections at d7 after intravitreal injections of NMDA or PBS. Retinas were immunolabeled for VEGFD (green). Nuclei were labeled with Hoechst (blue). GCL, ganglion cell layer. Scale bar, 20 μm. (F) qRT-PCR analysis of vegfd , vegfc , vegfa , and flt4 expression in brain ECs (b.END3) 24 h after 20 μM NMDA treatment. Unpaired t test. n = 3. vegfd , p = 0.1744; vegfc , p = 0.7598; vegfa , p = 0.8998; flt4 , p = 0.4457. (G) qRT-PCR analysis of vegfd , vegfc , vegfa , and flt4 expression in primary astrocytes 24 h after 20 μM NMDA treatment. Unpaired t test. n = 3. vegfd , p = 0.8716; vegfc , p = 0.4918; vegfa , p = 0.9902; flt4 , p = 0.9331. Bars represent mean ± SEM. Dots represent single values. **p

    Article Snippet: The following TaqMan gene expression assays were used in this study: Thy1 (Mm01174153_m1), nefl (Mm01315666_m1), ocld (Mm01282968_m1), cld5 (Mm00727012_m1), tjp1 (Mm00493699_s1), vegfd (Mm00438963_m1), vegfc (Mm01202432_m1), vegf (Mm01281449_m1), flt4 (Mm01292618_m1), flt1 (Mm00438980_m1), and kdr (Mm00440099_m1).

    Techniques: Expressing, Quantitative RT-PCR, Injection, Immunolabeling, Mouse Assay, Labeling

    Piperlongumine activates apoptosis and necroptosis, and inhibits cell migration and invasion A. RIP1-IP was performed on cell lysates from MPC cells treated with the indicated concentrations of PL at 21% and 1% O 2 for 24 hours and probed for RIP1 (top lane) and RIP3 (bottom lane). A representative image (n=3) is shown. B. MTT cells were treated with indicated concentrations of PL at 21% and 1% O 2 for 24 hours. Total cell lysates were analyzed by Western blot for cleaved PARP, cleaved caspase 3 and caspase 3. β-tubulin was used as a loading control. The representative image (n=3) is shown. C. 1.5 × 10 5 MPC cells were plated in the upper part of transwell chambers and allowed to migrate for 24 hours in the presence of 0, 1, and 5μM PL. The box and whiskers graph represents data from three independent experiments. D. 1.5 × 10 5 MPC cells were plated in the upper part of matrigel-coated transwell chambers and allowed to migrate for 24 hours in the presence of 0, 1, and 5μM PL. The box and whiskers graph represents data from three independent experiments. E. MPC cells were treated with indicated concentrations of PL for 24 hours. mRNA expression levels of Twist1, Vegfa, Mmp9, and Nanog were assessed by quantitative real-time PCR. The target gene transcript levels were normalized to Actb . A graph represents data from three independent experiments as mean +/− SEM. *P

    Journal: Oncotarget

    Article Title: Hypoxia potentiates the cytotoxic effect of piperlongumine in pheochromocytoma models

    doi: 10.18632/oncotarget.9643

    Figure Lengend Snippet: Piperlongumine activates apoptosis and necroptosis, and inhibits cell migration and invasion A. RIP1-IP was performed on cell lysates from MPC cells treated with the indicated concentrations of PL at 21% and 1% O 2 for 24 hours and probed for RIP1 (top lane) and RIP3 (bottom lane). A representative image (n=3) is shown. B. MTT cells were treated with indicated concentrations of PL at 21% and 1% O 2 for 24 hours. Total cell lysates were analyzed by Western blot for cleaved PARP, cleaved caspase 3 and caspase 3. β-tubulin was used as a loading control. The representative image (n=3) is shown. C. 1.5 × 10 5 MPC cells were plated in the upper part of transwell chambers and allowed to migrate for 24 hours in the presence of 0, 1, and 5μM PL. The box and whiskers graph represents data from three independent experiments. D. 1.5 × 10 5 MPC cells were plated in the upper part of matrigel-coated transwell chambers and allowed to migrate for 24 hours in the presence of 0, 1, and 5μM PL. The box and whiskers graph represents data from three independent experiments. E. MPC cells were treated with indicated concentrations of PL for 24 hours. mRNA expression levels of Twist1, Vegfa, Mmp9, and Nanog were assessed by quantitative real-time PCR. The target gene transcript levels were normalized to Actb . A graph represents data from three independent experiments as mean +/− SEM. *P

    Article Snippet: The probes included Mm00441533_g1, Mm03053917_g1, Mm01247357_m1, Mm01281449_m1, Mm04208233_g1, Mm00441531_m1, Mm02384862_g1.

    Techniques: Migration, MTT Assay, Western Blot, Expressing, Real-time Polymerase Chain Reaction

    Piperlongumine inhibits tumor growth and metastases formation and decreases expression levels of EMT and angiogenesis markers in vivo A. Nude female mice bearing MTT-Luc tumors were treated with 24 mg/kg/day PL (n=9) for 28 days. The control group was treated with vehicle (n=10). Tumor growth was assessed once a week. The graph shows a significant inhibition of tumor growth at all time points of treatment in the treated group (green) compared to control (red) animals. The graph represents data from 9 (treated) and 10 (vehicle) mice assessed at specific time points as mean +/− SEM. Statistical analysis was performed by Mann Whitney, U-test, at each time point. B. Representative tumors from control mice compared to the tumors from animals treated with PL. Scale bar: 1cm. C. Lungs were resected and subjected to bioluminescence imaging. The presence of lung metastases in treated mice was 46% lower than in control group. D. mRNA expression levels of Pou5f1, Twist1, Vegfa, Mmp9, and Nanog in tumors from both treated and control groups were assessed by quantitative real-time PCR. The target gene transcript levels were normalized to Actb . The box and whiskers graphs represent data from control (n=10) and treated (n=9) groups. *P

    Journal: Oncotarget

    Article Title: Hypoxia potentiates the cytotoxic effect of piperlongumine in pheochromocytoma models

    doi: 10.18632/oncotarget.9643

    Figure Lengend Snippet: Piperlongumine inhibits tumor growth and metastases formation and decreases expression levels of EMT and angiogenesis markers in vivo A. Nude female mice bearing MTT-Luc tumors were treated with 24 mg/kg/day PL (n=9) for 28 days. The control group was treated with vehicle (n=10). Tumor growth was assessed once a week. The graph shows a significant inhibition of tumor growth at all time points of treatment in the treated group (green) compared to control (red) animals. The graph represents data from 9 (treated) and 10 (vehicle) mice assessed at specific time points as mean +/− SEM. Statistical analysis was performed by Mann Whitney, U-test, at each time point. B. Representative tumors from control mice compared to the tumors from animals treated with PL. Scale bar: 1cm. C. Lungs were resected and subjected to bioluminescence imaging. The presence of lung metastases in treated mice was 46% lower than in control group. D. mRNA expression levels of Pou5f1, Twist1, Vegfa, Mmp9, and Nanog in tumors from both treated and control groups were assessed by quantitative real-time PCR. The target gene transcript levels were normalized to Actb . The box and whiskers graphs represent data from control (n=10) and treated (n=9) groups. *P

    Article Snippet: The probes included Mm00441533_g1, Mm03053917_g1, Mm01247357_m1, Mm01281449_m1, Mm04208233_g1, Mm00441531_m1, Mm02384862_g1.

    Techniques: Expressing, In Vivo, Mouse Assay, MTT Assay, Inhibition, MANN-WHITNEY, Imaging, Real-time Polymerase Chain Reaction

    A. Gating strategy for analyzing Sema3A expression among leukocytes, epithelial and endothelial cells in lung and dLN. B. Quantification of Sema3A positive cells in different cell populations in uninfected (n=4), and infected mice, at 3 days DPI (n=4) and 10 DPI (n=4) in lung (left panel) or dLN (right panel). Nrp1 Flox/Flox mice used. Data combined from two independent experiments. ns = not significant by two-way ANOVA. C. Quantification of Pkd1, Bnip3, Vegfa, Pdl1 and Sema3A mRNA level following 24 hour culture in 1% O2 chamber. D. Gating strategy (left) and representative histograms (right) analyzing Sema3A positive cell populations in B16.F10 tumors 11 days post-injection. Olfactory lobe is used as a positive control for Sema3A expression. E. Quantification of Sema3A positive cells in same experiment as in (D) in tumor, dLN and ndLN. F. Genomic organization of murine Sema3a gene, indicating where CRISPR guide RNA targets with red arrow (upper figure). MiSEQ sequence results for chosen Sema3A KO clone showing 4 base deletion in two alleles (46% of all reads), a frameshift in one allele (45% of all reads) and WT reads in 2% of all reads. G. RT-qPCR analysis show down- and up-regulation of Sema3A in Sema3A KO and OE cell lines, respectively. Normalized to Hprt. Experiment performed once. H. Intracellular staining shows no detectable expression of Sema3A in Sema3A KO cells, and expression in Sema3A OE cells, as expected. Experiment performed once, at low seeding density. I. Growth of WT, Sema3A KO and Sema3A OE B16.F10.Ova cell lines in normal, IFNγ or TNFα-rich media. Experiment performed once. Data indicate mean ± SD. ns = not significant by two-way ANOVA. Abbreviations: BEC, blood endothelial cells. dLN, draining lymph node. DPI, days post-infection. FRC, fibroblastic reticular cells. KO, knockout. LEC, lymphatic endothelial cells. ndLN, non-draining lymph node. OE, overexpressing. TME, tumor microenvironment. WT, wild-type.

    Journal: bioRxiv

    Article Title: Semaphorin 3A induces cytoskeletal paralysis in tumor-specific CD8+ T cells

    doi: 10.1101/849083

    Figure Lengend Snippet: A. Gating strategy for analyzing Sema3A expression among leukocytes, epithelial and endothelial cells in lung and dLN. B. Quantification of Sema3A positive cells in different cell populations in uninfected (n=4), and infected mice, at 3 days DPI (n=4) and 10 DPI (n=4) in lung (left panel) or dLN (right panel). Nrp1 Flox/Flox mice used. Data combined from two independent experiments. ns = not significant by two-way ANOVA. C. Quantification of Pkd1, Bnip3, Vegfa, Pdl1 and Sema3A mRNA level following 24 hour culture in 1% O2 chamber. D. Gating strategy (left) and representative histograms (right) analyzing Sema3A positive cell populations in B16.F10 tumors 11 days post-injection. Olfactory lobe is used as a positive control for Sema3A expression. E. Quantification of Sema3A positive cells in same experiment as in (D) in tumor, dLN and ndLN. F. Genomic organization of murine Sema3a gene, indicating where CRISPR guide RNA targets with red arrow (upper figure). MiSEQ sequence results for chosen Sema3A KO clone showing 4 base deletion in two alleles (46% of all reads), a frameshift in one allele (45% of all reads) and WT reads in 2% of all reads. G. RT-qPCR analysis show down- and up-regulation of Sema3A in Sema3A KO and OE cell lines, respectively. Normalized to Hprt. Experiment performed once. H. Intracellular staining shows no detectable expression of Sema3A in Sema3A KO cells, and expression in Sema3A OE cells, as expected. Experiment performed once, at low seeding density. I. Growth of WT, Sema3A KO and Sema3A OE B16.F10.Ova cell lines in normal, IFNγ or TNFα-rich media. Experiment performed once. Data indicate mean ± SD. ns = not significant by two-way ANOVA. Abbreviations: BEC, blood endothelial cells. dLN, draining lymph node. DPI, days post-infection. FRC, fibroblastic reticular cells. KO, knockout. LEC, lymphatic endothelial cells. ndLN, non-draining lymph node. OE, overexpressing. TME, tumor microenvironment. WT, wild-type.

    Article Snippet: The following TagMan probes were used: BNIP3 (Mm01275600-g1), HPRT (Mm03024075-m1), PDK1 (Mm00554300-m1), PDL1 (Mm00452054-m1), SEMA3A (Mm00436469-m1) and VEGFA (Mm00437306-m1).

    Techniques: Expressing, Infection, Mouse Assay, Injection, Positive Control, CRISPR, Sequencing, Quantitative RT-PCR, Staining, Knock-Out