Journal: PLoS ONE

Article Title: Combined Inhibition of Epidermal Growth Factor Receptor and Cyclooxygenase-2 Leads to Greater Anti-tumor Activity of Docetaxel in Advanced Prostate Cancer

doi: 10.1371/journal.pone.0076169

Figure Lengend Snippet: Cells grown in 1% FBS were exposed to 0.01 μmol/L docetaxel (D), 20 μmol/L gefitinib (G) or100 μmol/LNS-398 (N), alone or in combination, or with DMSO as control for 24 h. NF-KB, MMP-9 and VEGF mRNA (A and B) and protein levels (C and D) were measured as described in Materials and Methods. Values are reported as the mean±SD of three independent experiments.

Article Snippet: Antibodies against human VEGF and MMP-9 were obtained from R&D System and Bioworld, respectively.

Techniques: Control

Journal: Cells

Article Title: Effects of Human Deciduous Dental Pulp-Derived Mesenchymal Stem Cell-Derived Conditioned Medium on the Metabolism of HUVECs, Osteoblasts, and BMSCs

doi: 10.3390/cells11203222

Figure Lengend Snippet: Effect of VEGF-A in SHED-CM on the proliferative ability of HUVECs. The cells in the SHED-CM group exhibited significantly higher proliferative ability than those in the SHED-CM + anti-VEGF-A neutralizing antibody-supplemented (hereafter termed SHED-CM + anti-VEGF-A), VEGF-A, and control groups. Cells in the SHED-CM + anti-VEGF-A and VEGF-A groups showed significantly higher proliferative ability than those in the control group. ( n = 12, ** p < 0.01, * p < 0.05). SHED-CM, human deciduous dental pulp-derived mesenchymal stem cell-derived culture supernatant; VEGF-A, vascular endothelial growth factor-A; HUVECs, human umbilical vein endothelial cells.

Article Snippet: HUVECs were treated with SHED-CM in the presence or absence of 10 μg/mL anti-VEGF-A antibody (MAB293; R&D Systems, Minneapolis, MN, USA) in serum-free EBM-2 medium.

Techniques: Control, Derivative Assay

Journal: Cells

Article Title: Effects of Human Deciduous Dental Pulp-Derived Mesenchymal Stem Cell-Derived Conditioned Medium on the Metabolism of HUVECs, Osteoblasts, and BMSCs

doi: 10.3390/cells11203222

Figure Lengend Snippet: Effects of VEGF-A in SHED-CM on HUVEC lumen assembly. ( A ) Luminal view of HUVECs in the four groups. Lumen formation was observed more frequently in the SHED-CM- and VEGF-A-supplemented groups. Scale bar = 300 μm. ( B ) Vascular bifurcation in the four groups. ( C ) Vessel length in the four groups ( n = 4, ** p < 0.01). Black lines indicate vessel length; black arrows indicate joints. SHED-CM, human deciduous dental pulp-derived mesenchymal stem cell-derived culture supernatant; VEGF-A, vascular endothelial growth factor-A; HUVECs, human umbilical vein endothelial cells.

Article Snippet: HUVECs were treated with SHED-CM in the presence or absence of 10 μg/mL anti-VEGF-A antibody (MAB293; R&D Systems, Minneapolis, MN, USA) in serum-free EBM-2 medium.

Techniques: Derivative Assay

Journal: Chinese Medicine

Article Title: Shuxuening injection improves ventricular remodeling after myocardial infarction by indirectly promoting macrophage-mediated angiogenesis and lymphangiogenesis

doi: 10.1186/s13020-026-01352-w

Figure Lengend Snippet: SXNI promotes angiogenesis and lymphangiogenesis via regulating VEGF-A/VEGFR-2 and VEGF-C/VEGFR3 pathways. A Images of VEGF-A staining are depicted and IHC quantification of VEGF-A was conducted (n = 3). B ELISA analysis was used to determine serum VEGF-A levels in rats treated with SXNI at four weeks post-MI (n = 8). C Relative protein expression levels of VEGF-A and p-VEGFR2 in cardiac tissue were assessed (n = 3). D Images of VEGF-C staining are depicted and IHC quantification of VEGF-C is conducted (n = 3). E ELISA analysis of serum VEGF-C levels in rats treated with SXNI at four weeks post-MI is conducted (n = 8). F Relative VEGF-C and VEGFR3 protein expression levels in cardiac tissue are assessed (n = 3). Data are expressed as mean ± SD. # P < 0.05 , ## P < 0.01 , ### P < 0.001, vs the sham group; * P < 0.05, ** P < 0.01, *** P < 0.001, vs the model group; ns, no significant difference

Article Snippet: The following antibodies were used: monoclonal anti-CD31 antibody (1:200, AF6191, Affinity, Jiangsu, China) for staining microvessels, polyclonal anti-VEGF-A antibody (1:200, SC-7269, Santa Cruz Biotechnology, USA) for staining VEGF-A protein, monoclonal anti-LYVE-1 antibody (1:200, #67,538, CST, Boston, USA) for staining lymphatic vessels, and monoclonal anti-VEGF-C antibody (1:200, SC-374628, Santa Cruz Biotechnology, USA) for staining VEGF-C protein.

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Expressing

Journal: Chinese Medicine

Article Title: Shuxuening injection improves ventricular remodeling after myocardial infarction by indirectly promoting macrophage-mediated angiogenesis and lymphangiogenesis

doi: 10.1186/s13020-026-01352-w

Figure Lengend Snippet: SXNI promotes the protein release of angiogenesis factors and lymphangiogenesis factors in Raw264.7 cells. A Cell viability of Raw264.7 cells after SXNI treatment (n = 6). B , C Relative Vegf-a and Vegf-c mRNA levels in Raw264.7 cells after following 3 h or 9 h of SXNI exposure (n = 3 or 4). D , E VEGF-A and VEGF-C proteins released by Raw264.7 cells after being treated with SXNI for 24 h (n = 3 or 4). Data are expressed as mean ± SD. * P < 0.05, ** P < 0.01 , *** P < 0.001, vs the ctrl group; ns, no significant difference

Article Snippet: The following antibodies were used: monoclonal anti-CD31 antibody (1:200, AF6191, Affinity, Jiangsu, China) for staining microvessels, polyclonal anti-VEGF-A antibody (1:200, SC-7269, Santa Cruz Biotechnology, USA) for staining VEGF-A protein, monoclonal anti-LYVE-1 antibody (1:200, #67,538, CST, Boston, USA) for staining lymphatic vessels, and monoclonal anti-VEGF-C antibody (1:200, SC-374628, Santa Cruz Biotechnology, USA) for staining VEGF-C protein.

Techniques:

Journal: Chinese Medicine

Article Title: Shuxuening injection improves ventricular remodeling after myocardial infarction by indirectly promoting macrophage-mediated angiogenesis and lymphangiogenesis

doi: 10.1186/s13020-026-01352-w

Figure Lengend Snippet: The key active components in SXNI promote VEGF-A and VEGF-C proteins release in Raw264.7 cells. A Cell viability of Raw264.7 cells after ginkgolide A, ginkgolide B, rutin or quercetin 3-neohesperidoside treatment (n = 4). B , C Relative Vegf-a and Vegf-c mRNA levels in Raw264.7 cells after ginkgolide A, ginkgolide B, rutin or quercetin 3-neohesperidoside exposure (n = 3). D , E VEGF-A and VEGF-C proteins released by Raw264.7 cells after being treated with ginkgolide A, ginkgolide B, rutin or quercetin 3-neohesperidoside (n = 4). Data are expressed as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, versus the ctrl group; ns, no significant difference

Article Snippet: The following antibodies were used: monoclonal anti-CD31 antibody (1:200, AF6191, Affinity, Jiangsu, China) for staining microvessels, polyclonal anti-VEGF-A antibody (1:200, SC-7269, Santa Cruz Biotechnology, USA) for staining VEGF-A protein, monoclonal anti-LYVE-1 antibody (1:200, #67,538, CST, Boston, USA) for staining lymphatic vessels, and monoclonal anti-VEGF-C antibody (1:200, SC-374628, Santa Cruz Biotechnology, USA) for staining VEGF-C protein.

Techniques:

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Fibroblast growth factor 2 activation of stromal cell vascular endothelial growth factor expression and angiogenesis.

doi: 10.1038/labinvest.3780212

Figure Lengend Snippet: Figure 2. Temporal induction and regression of the angiogenic response to FGF-2–Matrigel implants. Neovascular basement membrane structures were evaluated in the Matrigel/skin stromal interface using antimouse type IV collagen immunostaining. Matrigel/skin cross-sections were immunostained with anti-type IV collagen to identify stromal neovasculature derived from control implants (MG1hep) or FGF-2 implants (MG1hep1FGF-2) isolated at 4, 7, and 14 days postimplantation. Dermis (D), stromal interface (I), and Matrigel (MG) structures are identified. Arrows depict new vessels in the stromal interface in the FGF-2–containing implant at 4 and 7 days. Note the prominent regression and Matrigel degradation at 14 days postimplantation in the FGF-2–containing implant.

Article Snippet: The capture antibody used was the chicken anti-VEGF polyclonal antibody coated at 5 mg/ml and the goat antimouse VEGF polyclonal antibody (R and D Systems, Minneapolis, Minnesota) used as detection antibody at 5 mg/ml, followed by horseradish peroxidase conjugated antigoat IgG at 0.5 mg/ml (Sigma).

Techniques: Membrane, Immunostaining, Derivative Assay, Control, Isolation

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Fibroblast growth factor 2 activation of stromal cell vascular endothelial growth factor expression and angiogenesis.

doi: 10.1038/labinvest.3780212

Figure Lengend Snippet: Figure 3. Correlation of VEGF and PECAM-1 immunostaining with FGF-2–containing Matrigel implants. A, Matrigel implants containing Matrigel alone (MG), Matrigel with heparin (MG1hep), and MG with heparin and FGF-2 (MG1hep1FGF-2) were evaluated for the expression of vascular endothelial growth factor (VEGF) and platelet-endothelial cell adhesion molecule 1 (PECAM-1) at 7 days postimplantation. Note the prominent detection of VEGF (anti-VEGF) in the FGF-2–containing implant stromal interface (arrows). Very little VEGF was detected in the heparin control and none in the Matrigel control. The detection of PECAM-1 (anti-PECAM-1) was also selectively restricted to the highly angiogenic FGF-2–induced stromal interface with both lumenal vascular capillary structures and individual cells invading into the Matrigel matrix (arrows). B, Higher power view of anti-PECAM-1 staining (blue signal) in Matrigel with heparin (MG 1 hep) and Matrigel with heparin and FGF-2 (MG1hep1FGF-2) (original magnification, 3900). Dermal, interface, and Matrigel regions indicated as D, I, and MG, respectively. Arrowheads show PECAM-1 positive capillaries in dermal regions and arrows show PECAM-1 positive vessels in the interface regions. C, Quantification of VEGF and PECAM-1 immunostaining in the FGF-2 and control Matrigel implants at Day 7 postimplantation. The number of positive cells per high-power field were averaged as described in “Materials and Methods.” Statistical significance of the FGF-2 implants compared with the Matrigel 1 heparin control was performed by Student’s t test for n 5 4. * p , 0.05; ** p , 0.001.

Article Snippet: The capture antibody used was the chicken anti-VEGF polyclonal antibody coated at 5 mg/ml and the goat antimouse VEGF polyclonal antibody (R and D Systems, Minneapolis, Minnesota) used as detection antibody at 5 mg/ml, followed by horseradish peroxidase conjugated antigoat IgG at 0.5 mg/ml (Sigma).

Techniques: Immunostaining, Expressing, Control, Staining

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Fibroblast growth factor 2 activation of stromal cell vascular endothelial growth factor expression and angiogenesis.

doi: 10.1038/labinvest.3780212

Figure Lengend Snippet: Figure 4. Left panel, VEGF and VEGF receptor-2/flk-1 expression in FGF-2–induced angiogenesis detected by in situ hybridization analysis. VEGF was detected with antisense probe (VEGF AS) and sense control (VEGF S) in FGF-2–containing Matrigel implants (7 days). Bright (B.F.) and dark inverted (D.F.) field photographs are presented. Arrows indicate strong focal cell staining. VEGF receptor-2/flk-1 was detected with antisense probe (FLK-1 AS) and sense control (FLK-1 S). Arrows indicate regions of positive staining. Dermis (D), stromal interface (I), and Matrigel (MG) are indicated. Right panel, Quantification of silver grains in the individual regions for VEGF antisense and sense and FLK-1 antisense and sense probes. The average of three fields in each region was measured and antisense compared with sense control for each region by Student’s t test. * p , 0.005.

Article Snippet: The capture antibody used was the chicken anti-VEGF polyclonal antibody coated at 5 mg/ml and the goat antimouse VEGF polyclonal antibody (R and D Systems, Minneapolis, Minnesota) used as detection antibody at 5 mg/ml, followed by horseradish peroxidase conjugated antigoat IgG at 0.5 mg/ml (Sigma).

Techniques: Expressing, In Situ Hybridization, Control, Staining

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Fibroblast growth factor 2 activation of stromal cell vascular endothelial growth factor expression and angiogenesis.

doi: 10.1038/labinvest.3780212

Figure Lengend Snippet: Figure 5. FGF-2 induces VEGF mRNA and protein in Balb/c 3T3 embryonic fibroblasts. A, Serum-restricted fibroblasts were treated with FGF-2 (10 ng/ml) (F), hypoxia (2% O2) (H), or untreated control (C). Northern blot analysis for VEGF (VEGF) and ribosome-associated protein (36B4) was performed (exposure time: 24 hours). B, VEGF protein secretion from control (2FGF-2) or FGF-2–treated (1FGF-2) Balb/c 3T3 fibroblasts over an 8-hour time period. Triplicate plate conditioned media samples were analyzed by ELISA and represented as average VEGF (ng/ml) 6 SEM. Statistically significant difference from untreated samples (* p , 0.05). C, Transient transfection and transcriptional activation of VEGF-promoter-luciferase reporter construct. Balb/c 3T3 cells were trans- fected in triplicate with either the VEGF promoter-luciferase construct or the pGL3 control vector. Postrecovery (18 hours), cells were treated with FGF-2 (FGF-2), hypoxia, or nothing (Control) for 8 hours. Cell lysates were analyzed for luciferase activity and subtracted from the pGL3 control levels. Data is presented as average light units per mg protein 6 SEM.

Article Snippet: The capture antibody used was the chicken anti-VEGF polyclonal antibody coated at 5 mg/ml and the goat antimouse VEGF polyclonal antibody (R and D Systems, Minneapolis, Minnesota) used as detection antibody at 5 mg/ml, followed by horseradish peroxidase conjugated antigoat IgG at 0.5 mg/ml (Sigma).

Techniques: Control, Northern Blot, Enzyme-linked Immunosorbent Assay, Transfection, Activation Assay, Luciferase, Construct, Plasmid Preparation, Activity Assay

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Fibroblast growth factor 2 activation of stromal cell vascular endothelial growth factor expression and angiogenesis.

doi: 10.1038/labinvest.3780212

Figure Lengend Snippet: Figure 6. FGF-2–induced VEGF expression is independent of cell adherence to attach- ment matrices. A, Serum-free Balb/c 3T3 cells were plated onto serum- (Plastic), Matrigel- (ECM), and fibronectin (Fn)-coated plates in the presence (FGF-2) or absence (control) of FGF-2 (10 ng/ml) for 8 hours. Phase photographs denote attachment to all three matrices and activation by FGF-2 (increase in rounded cell morphology). B, Top panel, Northern blot analysis for VEGF and 36B4 control from cells on the indicated matrices in the presence (F) or absence (C) of FGF-2 (10 ng/ml) for 8 hours. Bottom panel, Phosphorimage quantification of the VEGF mRNA signal normalized to the 36B4 control for Balb/c 3T3 cells plated and treated on each indicated matrix. Note the nearly identical FGF-2–induced VEGF mRNA levels for each matrix.

Article Snippet: The capture antibody used was the chicken anti-VEGF polyclonal antibody coated at 5 mg/ml and the goat antimouse VEGF polyclonal antibody (R and D Systems, Minneapolis, Minnesota) used as detection antibody at 5 mg/ml, followed by horseradish peroxidase conjugated antigoat IgG at 0.5 mg/ml (Sigma).

Techniques: Expressing, Control, Activation Assay, Northern Blot

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Fibroblast growth factor 2 activation of stromal cell vascular endothelial growth factor expression and angiogenesis.

doi: 10.1038/labinvest.3780212

Figure Lengend Snippet: Figure 7. Effect of systemic anti-VEGF antibodies on FGF-2–induced angiogenesis in vivo. A, Gross photographs of three representative Matrigel implants containing FGF-2 combined with systemic treatment with nonimmune chicken IgY (Control IgY) and anti-VEGF (Anti-VEGF IgY). Note the lack of associated skin vessel dilation and Matrigel coloration in the anti-VEGF group. B, Histologic evaluation of the stromal and angiogenic response induced by FGF-2 with control and anti-VEGF systemic treatments. Hematoxylin and eosin (H&E) and antimouse type IV collagen (collagen IV) are presented for control IgY (a and b) and anti-VEGF IgY (c and d). Matrigel (MG), stromal interface (I), and dermis (D) are indicated. Inset, Quantification of the angiogenic response in the control and anti-VEGF groups (n 5 10) is expressed as average vessels per field 6 SEM. Statistical difference from control IgG: * p , 0.001.

Article Snippet: The capture antibody used was the chicken anti-VEGF polyclonal antibody coated at 5 mg/ml and the goat antimouse VEGF polyclonal antibody (R and D Systems, Minneapolis, Minnesota) used as detection antibody at 5 mg/ml, followed by horseradish peroxidase conjugated antigoat IgG at 0.5 mg/ml (Sigma).

Techniques: In Vivo, Control