vascular endothelial growth factor  (R&D Systems)

Journal: Aging (Albany NY)

Article Title: Astrocyte-derived VEGF increases cerebral microvascular permeability under high salt conditions

doi: 10.18632/aging.103348

Figure Lengend Snippet: High salt activates astrocytes and up-regulates VEGF. ( A ) Representative images of double immunofluorescence staining for GFAP(red) and VEGF(green) in the brain specimens of rats; DAPI is stained in blue(n=5 per group). ( B , C ) Expression of GFAP and VEGF in the brain tissues from rats analyzed by western blotting and densitometry (n=5 per group). ( D ) Immunofluorescence staining of GFAP and AQP4 in red. ( E , F ) Expression of AQP4 and GFAP analyzed by western blotting in NaCl-treated astrocytes; * P <0.05, ** P <0.01, compared with NS. At least three separate experiments were conducted; means ± SD.

Article Snippet: Recombinant VEGF (CST, #5211), anti-VEGF neutralization antibody (R&D, #AF564) or SN50 (MCE, #213546-53-3) were added into the medium as indicated.

Techniques: Double Immunofluorescence Staining, Staining, Expressing, Western Blot, Immunofluorescence

Journal: Aging (Albany NY)

Article Title: Astrocyte-derived VEGF increases cerebral microvascular permeability under high salt conditions

doi: 10.18632/aging.103348

Figure Lengend Snippet: Increased expression of VEGF is mediated by NFκB/MMP-9 pathway. ( A , B ) HS increases protein levels of VEGF, MMP9 and p-p65. ( C ) Immunofluorescence staining of NaCl (40mM, 24 h)-induced p65 phosphorylation and nuclear translocation; ( D ) HS upregulates mRNA expression levels of VEGF. ( E ) SN50 antagonized NaCl-induced up-regulation of VEGF and MMP-9 protein expression. * P <0.05, ** P <0.01, compared with NS, n=3, means ± SD.

Article Snippet: Recombinant VEGF (CST, #5211), anti-VEGF neutralization antibody (R&D, #AF564) or SN50 (MCE, #213546-53-3) were added into the medium as indicated.

Techniques: Expressing, Immunofluorescence, Staining, Phospho-proteomics, Translocation Assay

Journal: Aging (Albany NY)

Article Title: Astrocyte-derived VEGF increases cerebral microvascular permeability under high salt conditions

doi: 10.18632/aging.103348

Figure Lengend Snippet: Astrocyte-derived VEGF mediates HS-induced BBB breakdown. ( A ) Representative double immunofluorescence staining of ZO-1 + bEnd.3 endothelium. Endothelial cells were cultured alone, or co-cultured with primary rats′ astrocytes, and treated with NS, HS, conditioned medium, or VEGF neutralizing antibody (NA). ( B , C ) Western blotting analysis of Occludin, Claudin-5, and ZO-1 in endothelial cells. ( D ) Permeability of tight junctions measured using NaF; * P <0.05, ** P <0.01, compared with EC+NS. At least three separate experiments were conducted; means ± SD.

Article Snippet: Recombinant VEGF (CST, #5211), anti-VEGF neutralization antibody (R&D, #AF564) or SN50 (MCE, #213546-53-3) were added into the medium as indicated.

Techniques: Derivative Assay, Double Immunofluorescence Staining, Cell Culture, Western Blot, Permeability

Journal: Aging (Albany NY)

Article Title: Astrocyte-derived VEGF increases cerebral microvascular permeability under high salt conditions

doi: 10.18632/aging.103348

Figure Lengend Snippet: Effect of VEGF on tight junction is mediated via ERK/eNOS. ( A , B ) Protein levels of ERK/pERK, eNOS/pSer 1177 , eNOS, and p38/p-p38 in endothelial cells; * P <0.05, ** P <0.01, compared with vehicle. At least three separate experiments were conducted; means ± SD.

Article Snippet: Recombinant VEGF (CST, #5211), anti-VEGF neutralization antibody (R&D, #AF564) or SN50 (MCE, #213546-53-3) were added into the medium as indicated.

Techniques:

Journal: Aging (Albany NY)

Article Title: Astrocyte-derived VEGF increases cerebral microvascular permeability under high salt conditions

doi: 10.18632/aging.103348

Figure Lengend Snippet: Increased cerebral microvascular permeability in VEGF hi/+ mice. ( A ) Schematic diagram of VEGF hi/+ mice generation. ( B ) Immunoblotting showing VEGF, Occludin, Claudin-5 and ZO-1expression in VEGF hi/+ and wildtype mice. ( C ) Cerebral cortical micro-vessel and dye leakage at 5 and 10 min after injection of Rhodamine B isothiocyanate-dextran; n=5, * P <0.05, ** P <0.01, compared with NSD. At least three separate experiments were conducted; means ± SD.

Article Snippet: Recombinant VEGF (CST, #5211), anti-VEGF neutralization antibody (R&D, #AF564) or SN50 (MCE, #213546-53-3) were added into the medium as indicated.

Techniques: Permeability, Western Blot, Injection

Journal: Aging (Albany NY)

Article Title: Astrocyte-derived VEGF increases cerebral microvascular permeability under high salt conditions

doi: 10.18632/aging.103348

Figure Lengend Snippet: Blocking VEGF attenuates disruption of tight junctions induced by HSD. ( A ) Representative images showing double immunofluorescence staining of ZO-1 in green and DAPI in the brain specimens of rats. ( B , C ) Expression of Occludin, Claudin-5 and ZO-1 determined by western blotting; n=5, * P <0.05, ** P <0.01, compared with NSD. At least three separate experiments were conducted; means ± SD.

Article Snippet: Recombinant VEGF (CST, #5211), anti-VEGF neutralization antibody (R&D, #AF564) or SN50 (MCE, #213546-53-3) were added into the medium as indicated.

Techniques: Blocking Assay, Disruption, Double Immunofluorescence Staining, Expressing, Western Blot

Journal: Scientific reports

Article Title: Extracellular Protein Fibulin-7 and Its C-Terminal Fragment Have In Vivo Antiangiogenic Activity.

doi: 10.1038/s41598-018-36182-w

Figure Lengend Snippet: Figure 2. Fbln7 and Fbln7-C bind to VEGFR2, but not to VEGFR1 or VEGF. Binding between Fbln7-FL/ Fbln7-C and (A) VEGF, (B) VEGFR1, or (C) VEGFR2 by ELISA. FN: Fibronectin, FL: Fbln7-FL, C: Fbln7-C, V: VEGF. **P < 0.01. (D) Binding between Fbln7-C and VEGFR2 by pull-down assays. (E) Binding between Fbln7-C and VEGFR2 at various protein ratios: (1) VEGFR2:Fbln7-C = 1:1; (2) VEGFR2:Fbln7-C = 1:10; (3) VEGFR2:Fbln7-C = 1:20. All samples included the VEGFR2 protein. Each row is a different gel, but every gel was loaded with the same sample in equal volumes.

Article Snippet: Recombinant vascular endothelial growth factor (VEGF) 164 (493-MV-005/CF), recombinant VEGFR2/Flk-1 Fc Chimera (443-KD-050/CF), and recombinant VEGFR1 Fc Chimera (471-F1-100) were purchased from R&D Systems (Minneapolis, MN).

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay

Journal: Scientific reports

Article Title: Extracellular Protein Fibulin-7 and Its C-Terminal Fragment Have In Vivo Antiangiogenic Activity.

doi: 10.1038/s41598-018-36182-w

Figure Lengend Snippet: Figure 3. Fbln7-C inhibits VEGFR2 phosphorylation and ERK phosphorylation in the VEGF-VEGFRs signaling pathway. (A,B) VEGFR2 Tyr1175, and (C,D) ERK phosphorylation levels with Fbln7-C (100 µg/ml) pretreatment and after VEGF (5 ng/ml) stimulation were confirmed by western blotting. C: control, F7C: Fbln7-C *P < 0.05. All samples included the VEGFR2 protein. Each row is a different gel, but every gel was loaded with the same sample in equal volumes.

Article Snippet: Recombinant vascular endothelial growth factor (VEGF) 164 (493-MV-005/CF), recombinant VEGFR2/Flk-1 Fc Chimera (443-KD-050/CF), and recombinant VEGFR1 Fc Chimera (471-F1-100) were purchased from R&D Systems (Minneapolis, MN).

Techniques: Phospho-proteomics, Western Blot, Control

Journal: Scientific reports

Article Title: Extracellular Protein Fibulin-7 and Its C-Terminal Fragment Have In Vivo Antiangiogenic Activity.

doi: 10.1038/s41598-018-36182-w

Figure Lengend Snippet: Figure 5. Fbln7-C affects focal adhesion area and actin filaments to inhibit cell motility. (A–C) Cell motility on fibronectin or Fbln7-C-coated dishes stimulated with VEGF and analyzed using time-lapse imaging. Cell motility is evaluated by (A) cell velocity, (B) distance, (C) and persistence. N = 3 n > 150, ****P < 0.0001, **P < 0.01, *P < 0.01. (D–I) Cell morphology differences between cells on fibronectin and Fbln7-C-coated dishes evaluated by staining for focal adhesion sites (paxillin; red) and actin filaments (phalloidin; green). (D) Immunofluorescence staining. Cell shape is evaluated by (E) shape factor and (F) number of lamellipodia. Focal adhesion area is evaluated by (G) average focal adhesion area, (H) focal adhesion number and (I) cell area. FN: Fibronectin, F7C:Fibulin7-C, scale bar: 100 μm, N = 3 n > 80, ****P < 0.0001, **P < 0.01, *P < 0.01.

Article Snippet: Recombinant vascular endothelial growth factor (VEGF) 164 (493-MV-005/CF), recombinant VEGFR2/Flk-1 Fc Chimera (443-KD-050/CF), and recombinant VEGFR1 Fc Chimera (471-F1-100) were purchased from R&D Systems (Minneapolis, MN).

Techniques: Imaging, Staining, Immunofluorescence