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Rhvegf c

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RhVEGF-C is a recombinant human protein that functions as a vascular endothelial growth factor. It is used for research purposes.

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Lab products found in correlation

Lsrii flow cytometry, by BD (2 mentions) Cd45.1 b6 sjl ptprca pepcb boyj mice, by Jackson ImmunoResearch (2 mentions) Collagenase d, by Merck Group (2 mentions) Magnetic microbead based positive selection kit for cd11c cells, by Miltenyi Biotec (2 mentions) Erk1 2, by Cell Signaling Technology (1 mentions) P erk1 2, by Cell Signaling Technology (1 mentions) Nitrocellulose membrane, by GE Healthcare (1 mentions) Vegfr 3, by Merck Group (1 mentions) Gadph, by Merck Group (1 mentions) Rhvegfa, by R&D Systems (1 mentions) P vegfr2, by Cell Signaling Technology (1 mentions) Vegfr2, by Cell Signaling Technology (1 mentions) Raw264, by American Type Culture Collection (1 mentions) α minimal essential medium, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Hepes, by Merck Group (1 mentions) Dimethylformamide, by Merck Group (1 mentions) Human macrophage colony stimulating factor m csf, by R&D Systems (1 mentions)

Close spelling variants of this product

RhVEGF (14 citations)

7 protocols using rhvegf c

1

Modulation of VEGFR Signaling by AD0157

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Serum-starved subconfluent LEC cultures were incubated in the presence or absence of AD0157 for 2 h and then challenged for 30 additional minutes with rhVEGF-C (400 ng/mL), rhVEGF-C156S (500 ng/mL; R&D Systems), or rhVEGF-A (100 ng/mL; R&D Systems). Cell lysates were analyzed by SDS-PAGE, electrotransferred to nitrocellulose membranes (GE Healthcare Life Sciences), and blots were probed with primary antibodies against pVEGFR-3 (1/1000; Cell Application), VEGFR-3 (1/1000; Millipore), pVEGFR-2, VEGFR-2, pERK1/2, ERK1/2, pAkt, Akt, (1/1000; Cell Signaling), and GADPH (1/2000; Millipore). Detection was carried out with chemiluminescence system ECL Western Blotting Substrate (Pierce), and bands were quantified and expressed as phosphorylated protein/total protein ratio.
García-Caballero M., Paupert J., Blacher S., Van de Velde M., Quesada A.R., Medina M.A, & Noël A. (2017). Targeting VEGFR-3/-2 signaling pathways with AD0157: a potential strategy against tumor-associated lymphangiogenesis and lymphatic metastases. Journal of Hematology & Oncology, 10, 122.
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2

Intradermal VEGF-C Modulates Dendritic Cell Trafficking

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1µl of rhVEGF-C (0.1mg/ml, R&D, Minneapolis, MN) was injected intradermally with a Hamilton syringe into the base of the ear in obese and lean animals every day for 7 days prior to induction of contact hypersensitivity and for 3 days post-challenge for a total of 10 days (Szuba et al., 2002 (link)). Control animals were injected with 1µl of sterile PBS. To quantify dendritic cell (DC) trafficking, we used a modification of previously reported methods (Lim et al., 2009 (link)). Briefly, the spleens of CD45.1 B6.SJL-Ptprca Pepcb/BoyJ mice (Jackson Laboratories) were harvested and digested with collagenase D (Sigma-Aldrich). CD45.1+ DCs were enriched using a magnetic microbead-based positive selection kit for CD11c+ cells (Miltenyi Biotech, Gladbach, Germany). Isolated cells were resuspended in PBS and injected into the apex of the right ear (106 cells per injection), and 18 hours later the right cervical lymph nodes were harvested and analyzed for migrating DCs (CD45.1+/CD11c+/MHC-IIhigh) using flow cytometry (LSRII;BD Biosciences, San Jose, CA) and FlowJo software (Tree Star, Ashland, OR). Each experiment was repeated with 4–5 animals per group.
Savetsky I.L., Albano N.J., Cuzzone D.A., Gardenier J.C., Torrisi J.S., García Nores G.D., Nitti M.D., Hespe G.E., Nelson T.S., Kataru R.P., Dixon J.B, & Mehrara B.J. (2015). Lymphatic function regulates contact hypersensitivity dermatitis in obesity. The Journal of investigative dermatology, 135(11), 2742-2752.
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3

Osteoclast Differentiation from Mouse Cells

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The mouse bone marrow cells and the macrophage/pre-osteoclast cell line RAW264.7 (purchased from ATCC, Manassas, VA, USA) were maintained in α-minimal essential medium (MEM; Life Technologies, Grand Island, NY, USA), supplemented with 10% fetal bovine serum (FBS; CSL Biosciences, VIC, Australia), 0.005% penicillin (10,000 U/ml)/streptomycin (10,000 U/ml) (Life Technologies, USA), L-glutamine (Life Technologies) and HEPES (Sigma-Aldrich, USA). Cells were cultured at 37°C with 5% CO2 and 100% humidity. Recombinant soluble murine receptor activator of nuclear factor κB ligand (RANKL) was purchased from the Oriental Yeast Co. (Tokyo, Japan). For tartrate resistant acid phosphatase (TRAP) staining, naphthol AMX phosphate, fast red violet LB Salt F-1625, and dimethylformamide were purchased from Sigma-Aldrich (St Louis, MO, USA). The human macrophage-colony stimulating factor (M-CSF) was purchased from R&D Systems (Minneapolis, MN, USA). The rhVEGF-C and rhVEGF-D were purchased from R&D Systems.
Zhu Y., Wu Y., Liang Y., Tan W., Liu Z, & Xiao J. (2015). Regulation of expression level of fms-like tyrosine kinase-4 is related to osteoclast differentiation. Archives of Medical Science : AMS, 12(3), 502-506.
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4

Intradermal VEGF-C Modulates Dendritic Cell Trafficking

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1µl of rhVEGF-C (0.1mg/ml, R&D, Minneapolis, MN) was injected intradermally with a Hamilton syringe into the base of the ear in obese and lean animals every day for 7 days prior to induction of contact hypersensitivity and for 3 days post-challenge for a total of 10 days (Szuba et al., 2002 (link)). Control animals were injected with 1µl of sterile PBS. To quantify dendritic cell (DC) trafficking, we used a modification of previously reported methods (Lim et al., 2009 (link)). Briefly, the spleens of CD45.1 B6.SJL-Ptprca Pepcb/BoyJ mice (Jackson Laboratories) were harvested and digested with collagenase D (Sigma-Aldrich). CD45.1+ DCs were enriched using a magnetic microbead-based positive selection kit for CD11c+ cells (Miltenyi Biotech, Gladbach, Germany). Isolated cells were resuspended in PBS and injected into the apex of the right ear (106 cells per injection), and 18 hours later the right cervical lymph nodes were harvested and analyzed for migrating DCs (CD45.1+/CD11c+/MHC-IIhigh) using flow cytometry (LSRII;BD Biosciences, San Jose, CA) and FlowJo software (Tree Star, Ashland, OR). Each experiment was repeated with 4–5 animals per group.
Savetsky I.L., Albano N.J., Cuzzone D.A., Gardenier J.C., Torrisi J.S., García Nores G.D., Nitti M.D., Hespe G.E., Nelson T.S., Kataru R.P., Dixon J.B, & Mehrara B.J. (2015). Lymphatic function regulates contact hypersensitivity dermatitis in obesity. The Journal of investigative dermatology, 135(11), 2742-2752.
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5

Quantitative Migration and Invasion Assay

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The migratory activity was assessed using the scratch assay. Cells in a confluent monolayer were scratched and supplied with complete medium containing 2% FBS, in the absence or presence of AD0157. Wounded areas were photographed at 0 and 48 h of incubation. Mitomycin C (0.1 μg/ml; Sigma-Aldrich) was added to inhibit cellular proliferation. The percentage of recovered area was determined by image analysis (NIH Image 1.6 software). For invasion assays, LECs (1.5 × 105) in EBM-2 medium containing 0.5% FBS, with or without AD0157, were seeded in the upper compartment of Transwell inserts (Corning) coated with 0.2% gelatin. The lower compartment was filled with EBM-2 containing 1% FBS and rhVEGF-C (400 ng/ml; R&D Systems). After 48 h, cells were fixed, stained with Giemsa solution (Millipore), and filters were visualized and quantified.
García-Caballero M., Paupert J., Blacher S., Van de Velde M., Quesada A.R., Medina M.A, & Noël A. (2017). Targeting VEGFR-3/-2 signaling pathways with AD0157: a potential strategy against tumor-associated lymphangiogenesis and lymphatic metastases. Journal of Hematology & Oncology, 10, 122.
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6

Lymphatic Endothelial Cell Proliferation Assay

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Human lymphatic microvascular endothelial cells (HMVEC-dLy-Neo: CC-2812) were obtained from TaKaRa (Shiga, Japan) and cultured in EGM-2-MV BulletKit medium at 37°C and 5% humidified CO2. The HMVECs were seeded into a 96-well plate at 3 × 103 cells/wells and were allowed to adhere overnight. The culture medium was then replaced and the cells were cultured with PGE2 (1 or 10 nM), an EP1-4 receptor agonist, or rhVEGF-C (10 μg/mL; R&D Systems, Minneapolis, MN, USA) for 48 h. HMVEC proliferation was measured using a Cell Counting Kit-8 assay (Dojindo, Kumamoto, Japan), according to the manufacturer's instructions.
Hosono K., Isonaka R., Kawakami T., Narumiya S, & Majima M. (2016). Signaling of Prostaglandin E Receptors, EP3 and EP4 Facilitates Wound Healing and Lymphangiogenesis with Enhanced Recruitment of M2 Macrophages in Mice. PLoS ONE, 11(10), e0162532.
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7

Lymphatic Vessel Response to VEGF-C in UV-Induced Inflammation

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WT mice were randomized to receive intradermal injections of recombinant human VEGF-C (rhVEGF-C; R&D Systems; Minneapolis, MN) or an equal volume of PBS on days 0, 2, 4, 6, 8, and 11 (see additional details below). These mice were compared to LECPTEN mice injected with vehicle at the same time points. After the first injection, each mouse was placed in an opaque-walled cage, where it could roam freely. Ultraviolet radiation (UVR) light was delivered by four FS40T12/UVB/BP bulbs (Light Sources, Inc.; Orange, CT) placed on top of the cage for 5:02 minutes to deliver a total dose of 1000 J/m2, which was quantified by UVA/B Light Meter 850009 (Sper Scientific; Scottsdale, AZ). Ear inflammation was subsequently quantified by measuring skin thickness with digital calipers on days 3, 7, and 14. Ear samples were also collected, fixed, and processed to assess lymphatic vessels and inflammatory cells as described below.
Kataru R.P., Baik J.E., Park H.J., Ly C.L., Shin J., Schwartz N., Lu T.T., Ortega S, & Mehrara B.J. (2021). Lymphatic-specific intracellular modulation of receptor tyrosine kinase signaling improves lymphatic growth and function. Science signaling, 14(695), eabc0836.
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