Journal: PLoS ONE

Article Title: Increased Carotid Intima-Media Thickness and Reduced Distensibility in Human Class III Obesity: Independent and Differential Influences of Adiposity and Blood Pressure on the Vasculature

doi: 10.1371/journal.pone.0053972

Figure Lengend Snippet: Representative flow cytometric density plots demonstrating the gating protocol used to identify angiogenic cells: A) PBMCs stained with PE-conjugated and APC-conjugated mouse IgG (isotype controls) for non-specific fluorescent signals; B) PBMCs stained with antibodies against human CD133 (AC133) (PE-conjugated) and VEGFR-2 (KDR) (APC-conjugated) for AC133 + /KDR + PBMCs; A1, B1) FSC-SSC density dot plots of Ficoll-isolated PBMCs and R1 was gated for monocytes; A2, B2) FL2 (PE)-FL4 (APC) density dot plots of R1-gated monocytes. Angiogenic cells = cells in B2 upper-right quadrant – cells in A2 upper-right quadrant.

Article Snippet: In brief, 100 µl of PBMCs (1.5×10 7 /ml) was incubated with Fc-γ receptor blocking agent followed by 30 min incubation on ice with antibodies against human CD133 (PE-conjugated, Miltenyi Biotec, Germany) and VEGFR-2 (KDR) (APC-conjugated, R&D Systems, USA).

Techniques: Staining, Isolation

Journal: mAbs

Article Title: Analytical and functional similarity of Amgen biosimilar ABP 215 to bevacizumab

doi: 10.1080/19420862.2018.1452580

Figure Lengend Snippet: Inhibition of VEGFR-2 RTK autophosphorylation by ABP 215, bevacizumab (US), and bevacizumab (EU).

Article Snippet: VEGFR-2 was captured from the lysate onto streptavidin-coated Mesoscale Discovery (MSD, Cat. No. L15SA) plates using a biotinylated antibody against the extracellular portion of VEGFR-2 (R&D Systems, Cat. No. 05–321).

Techniques: Inhibition

Journal: PLoS ONE

Article Title: Vascular Endothelial Growth Factor Receptor-3 Directly Interacts with Phosphatidylinositol 3-Kinase to Regulate Lymphangiogenesis

doi: 10.1371/journal.pone.0039558

Figure Lengend Snippet: A , Prostatic LEC tube length at 4.5 hours post VEGF-C treatment was quantified using ImageJ. VEGF-C significantly increased the number of tubes formed compared to vehicle control. Data expressed as mean±s.e.m., n = 3, *** P <0.001 using One-way ANOVA, Bonferroni post-analysis. B , Western blotting analysis of VEGFR-2 and VEGFR-3 expression in lung, neonatal dermis and prostate LECs. β-tubulin was used as a loading control.

Article Snippet: The other antibodies used were: anti-phospho PI3K p85 (Tyr458/p55 Tyr199; Cat. no. 4228; Millipore, MA) and total PI3K p85 (Cat. no. 06–19; Millipore); mouse anti-human CD31 (Clone: JC70A, Cat. no. M0823; DAKO, Denmark); mouse anti-human podoplanin (Clone: D2-40, Cat. no. 730-23; Signet Laboratories, MA); mouse anti-human VEGFR-2 (Cat. no. AF357; R&D Systems, MN), goat anti-human VEGFR-3 (Cat. no. AF349; R&D Systems); mouse anti-human pan-actin (Clone: Ab-5, NeoMarkers, CA); Alexa Fluor 488- and 568-conjugated secondary antibodies (Invitrogen, CA), and IRDye700DX conjugated anti-mouse IgG and IRDye800CW conjugated anti-rabbit IgG (Rockland Immunochemicals, PA).

Techniques: Western Blot, Expressing

Journal: PLoS ONE

Article Title: Vascular Endothelial Growth Factor Receptor-3 Directly Interacts with Phosphatidylinositol 3-Kinase to Regulate Lymphangiogenesis

doi: 10.1371/journal.pone.0039558

Figure Lengend Snippet: . A , Western blotting analysis of phosphorylated Akt (S473) in prostatic LECs following a 15 minute ligand stimulation: VEGF-A (100 ng/ml), VEGF-C (100 ng/ml), VEGF-C156S (250 ng/ml), VEGF-D (250 ng/ml) and VEGF-E (100 ng/ml). B , Time course for Akt phosphorylation (S473) in LECs after VEGF-C (100 ng/ml) stimulation. C , Concentration-dependent phosphorylation of LEC Akt (S473) following 15 minute stimulation with VEGF-C or VEGF-C156S. D , Effect of inhibition of VEGFR-3 (hF4-3C5), VEGFR-2 (IMC-1121b) or VEGFR-1 (IMC-18F1) on LEC Akt (S473) phosphorylation in response to VEGF-C (100 ng/ml). E , Effect of inhibition of PI3K (AS252424, LY294002) or Raf/MEK (PD98059) on Akt (S473) phosphorylation in LECs in response to VEGF-C (100 ng/ml). Serum-free vehicle treated LEC lysate is indicated by ‘0′ in all blots. n = 3. Densitometry analysis is shown for each blot either italicized or graphed; where integrated intensity of phosphorylated molecules was firstly compared to that of the total target protein for each sample, and then expressed as fold increase in integrated density compared to VEGF-C treated control samples. P value calculated using one-way ANOVA. For panels A-B data is compared to time-point zero; panels D-E data is compared to VEGF-C treated control, except where indicated for serum free/VEGF-C treatment comparison. Columns: mean; bars: s.e.m.; P<0.05 (*), P<0.01 (**); P<0.001 (***).

Article Snippet: The other antibodies used were: anti-phospho PI3K p85 (Tyr458/p55 Tyr199; Cat. no. 4228; Millipore, MA) and total PI3K p85 (Cat. no. 06–19; Millipore); mouse anti-human CD31 (Clone: JC70A, Cat. no. M0823; DAKO, Denmark); mouse anti-human podoplanin (Clone: D2-40, Cat. no. 730-23; Signet Laboratories, MA); mouse anti-human VEGFR-2 (Cat. no. AF357; R&D Systems, MN), goat anti-human VEGFR-3 (Cat. no. AF349; R&D Systems); mouse anti-human pan-actin (Clone: Ab-5, NeoMarkers, CA); Alexa Fluor 488- and 568-conjugated secondary antibodies (Invitrogen, CA), and IRDye700DX conjugated anti-mouse IgG and IRDye800CW conjugated anti-rabbit IgG (Rockland Immunochemicals, PA).

Techniques: Western Blot, Concentration Assay, Inhibition

Journal: PLoS ONE

Article Title: Vascular Endothelial Growth Factor Receptor-3 Directly Interacts with Phosphatidylinositol 3-Kinase to Regulate Lymphangiogenesis

doi: 10.1371/journal.pone.0039558

Figure Lengend Snippet: Western blotting analysis of phosphorylated P70S6K ( A, top left ) and eNOS (S1177) ( B, top left ) in prostatic LECs following 15 minute stimulation with ligand: VEGF-C (100 ng/ml), VEGF-A (100 ng/ml), VEGF-C156S (250 ng/ml), VEGF-D (250 ng/ml) and VEGF-E (100 ng/ml). Time- and concentration-dependent phosphorylation of P70S6K ( A, top right ), eNOS (S1177) ( B, top right ), and PLCγ1 (Tyr783) ( C, top left ) in LECs in response to VEGF-C. The effect of inhibition of VEGFR-3 (hF4-3C5), VEGFR-2 (IMC-1121b) or VEGFR-1 (IMC-18F1) on phosphorylation of P70S6K ( A, bottom left ), eNOS (S1177) ( B, bottom left ); and VEGFR-3 (hF4-3C5) on PLCγ1 (Tyr783) ( C, bottom right ) on LEC response to VEGF-C (100 ng/ml). The effect of AS252424, LY294002, PD98059, and U-73122 on phosphorylation of P70S6K ( A, top right ), eNOS (S1177) ( B, bottom right ), and PLCγ1 (Tyr783) ( C, bottom right ) on LECs in response to VEGF-C. The effect of VEGF-C on phosphorylation of PLCγ2 (Tyr759, Tyr1217) in LECs ( C, bottom left ). Control serum-free vehicle treated LEC lysate is indicated by ‘0′ in all blots. n = 3. Densitometry analysis is shown under each blot in italics; where integrated intensity of phosphorylated molecules was firstly compared to that of the total target protein for each sample, and then expressed as fold change in integrated density compared to either serum-free (A, top left and right panels ; B, top left and right panels ; C, top and bottom left panels ) or VEGF-C treated control samples (A, bottom left and right panels ; B, bottom left and right panels ; C, bottom and bottom left panels ).

Article Snippet: The other antibodies used were: anti-phospho PI3K p85 (Tyr458/p55 Tyr199; Cat. no. 4228; Millipore, MA) and total PI3K p85 (Cat. no. 06–19; Millipore); mouse anti-human CD31 (Clone: JC70A, Cat. no. M0823; DAKO, Denmark); mouse anti-human podoplanin (Clone: D2-40, Cat. no. 730-23; Signet Laboratories, MA); mouse anti-human VEGFR-2 (Cat. no. AF357; R&D Systems, MN), goat anti-human VEGFR-3 (Cat. no. AF349; R&D Systems); mouse anti-human pan-actin (Clone: Ab-5, NeoMarkers, CA); Alexa Fluor 488- and 568-conjugated secondary antibodies (Invitrogen, CA), and IRDye700DX conjugated anti-mouse IgG and IRDye800CW conjugated anti-rabbit IgG (Rockland Immunochemicals, PA).

Techniques: Western Blot, Concentration Assay, Inhibition

Journal: PLoS ONE

Article Title: Differential Expression of Vegfr-2 and Its Soluble Form in Preeclampsia

doi: 10.1371/journal.pone.0033475

Figure Lengend Snippet: A : Western blot analysis of VEGFR-1 and VEGFR-2 from gestational age-matched normal pregnant women (NP, 1–5) and preeclamptic women (PE, 6–10). B : Maternal plasma concentration of VEGFR1, VEGFR2 and Eng (mean±SE, pg/ml) at inclusion (Incl), delivery (day 0, D 0) and during post partum (day 1 to day 5, D 1 to D 5). PE: severe preeclampsia (plain line), NP: normal pregnancies (dashed line).

Article Snippet: After being washed, the membranes were incubated overnight at 4°C with either the VEGFR-2 antibody (AF357, diluted 1∶500, R&D Systems) or the VEGFR-1 antibody (AF321, diluted 1∶500, R&D Systems) in TBS-T containing 1% BSA.

Techniques: Western Blot, Concentration Assay

Journal: PLoS ONE

Article Title: Differential Expression of Vegfr-2 and Its Soluble Form in Preeclampsia

doi: 10.1371/journal.pone.0033475

Figure Lengend Snippet: A : Schematic representation of pre-mRNA exon-intron structure of VEGFR-2 (central shadowed box not at scale) with primers used to discriminate membrane bound VEGFR-2 mRNA (upper box, exon 13-exon 16; 634 bp) and soluble VEGFR-2 mRNA (lower box, exon 13-intron 13; 278 bp). B : Relative mRNA levels are expressed as arbitrary units (A.U.). NP: normal pregnancies, PE: severe preeclampsia. ** P <0.001 compared to NP group; ns: non-significant compared to NP group.

Article Snippet: After being washed, the membranes were incubated overnight at 4°C with either the VEGFR-2 antibody (AF357, diluted 1∶500, R&D Systems) or the VEGFR-1 antibody (AF321, diluted 1∶500, R&D Systems) in TBS-T containing 1% BSA.

Techniques:

Journal: PLoS ONE

Article Title: Differential Expression of Vegfr-2 and Its Soluble Form in Preeclampsia

doi: 10.1371/journal.pone.0033475

Figure Lengend Snippet: Placental villi from a normal pregnant women (A, C, E and F) and from a preeclamptic women (B–D): VEGFR-1 is expressed in the cytotrophoblasts, syncytiotrophoblasts (arrow) and also in some endothelial cells (white arrow head). VEGFR-2 is mainly localized in vascular endothelial cells (arrows) in placentas of both groups as identified by serial section immunostaining with CD31.

Article Snippet: After being washed, the membranes were incubated overnight at 4°C with either the VEGFR-2 antibody (AF357, diluted 1∶500, R&D Systems) or the VEGFR-1 antibody (AF321, diluted 1∶500, R&D Systems) in TBS-T containing 1% BSA.

Techniques: Immunostaining

Journal: BMC Cancer

Article Title: Anti-lymphangiogenic properties of mTOR inhibitors in head and neck squamous cell carcinoma experimental models

doi: 10.1186/1471-2407-13-320

Figure Lengend Snippet: The level of soluble VEGFR-2 was evaluated in serum of SCID mice that were sacrificed on day 21 after injection of the OSC-19 cells. Rapamycin treatment significantly increased the level of soluble VEGFR-2 in serum of SCID mice (p = 0.0001; t-test). Serum samples from 9 control mice and 10 rapamycin-treated mice were evaluated.

Article Snippet: The effects of rapamycin treatment on serum levels of soluble VEGFR-2 in mouse serum samples were determined using a mouse VEGFR-2 ELISA kit (R&D Systems, Minneapolis, MN) according to manufacturer's instructions.

Techniques: Injection

Journal: Journal of Cancer

Article Title: Evaluation of Detection Methods and Values of Circulating Vascular Endothelial Growth Factor in Lung Cancer

doi: 10.7150/jca.22020

Figure Lengend Snippet: The information of ten studies detecting VEGF level in serum.

Article Snippet: Tony Mok , Asia Caucasian , 303 , Protective Multicenter Randomized , ELISA(R&D Systems Minneapolis, MN) , Microplate reader (Bio-Tek Elx 800) , VEGF,VEGFR-1,VEGFR-2,bFGF,ICAM,PlGF, E-selection , 0, every 6week , IIIB-IV /NSCLC , Chemotherapy.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Journal of Cancer

Article Title: Evaluation of Detection Methods and Values of Circulating Vascular Endothelial Growth Factor in Lung Cancer

doi: 10.7150/jca.22020

Figure Lengend Snippet: The information of sixteen studies detecting VEGF level in plasma.

Article Snippet: Tony Mok , Asia Caucasian , 303 , Protective Multicenter Randomized , ELISA(R&D Systems Minneapolis, MN) , Microplate reader (Bio-Tek Elx 800) , VEGF,VEGFR-1,VEGFR-2,bFGF,ICAM,PlGF, E-selection , 0, every 6week , IIIB-IV /NSCLC , Chemotherapy.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Journal of Cancer

Article Title: Evaluation of Detection Methods and Values of Circulating Vascular Endothelial Growth Factor in Lung Cancer

doi: 10.7150/jca.22020

Figure Lengend Snippet: The information of studies detecting VEGF level in sputum, MPE, and EBC.

Article Snippet: Tony Mok , Asia Caucasian , 303 , Protective Multicenter Randomized , ELISA(R&D Systems Minneapolis, MN) , Microplate reader (Bio-Tek Elx 800) , VEGF,VEGFR-1,VEGFR-2,bFGF,ICAM,PlGF, E-selection , 0, every 6week , IIIB-IV /NSCLC , Chemotherapy.

Techniques: