anti vegf164 neutralizing antibody  (R&D Systems)

Journal: Scientific Reports

Article Title: A novel small molecule ameliorates ocular neovascularisation and synergises with anti-VEGF therapy

doi: 10.1038/srep25509

Figure Lengend Snippet: ( a ) Representative OCT images obtained 7 days post-laser, showing CNV lesions of vehicle (left), anti-VEGF164 (middle), and 1 μM SH-11037 (right). Scale bars = 100 μm. ( b ) Representative images from fluorescein angiography 14 days post-laser. ( c ) Representative images from confocal microscopy for agglutinin-stained CNV lesions 14 days post-laser, scale bars = 50 μm. ( d ) Quantification of CNV lesion volumes from OCT images at day 7 using ellipsoid volume measurement . ( e ) Quantification of CNV lesion volumes from Z-stack images at day 14 using ImageJ software. ** P < 0.01, one-way ANOVA, Tukey’s post hoc tests, ns; non significant. Mean ± s.e.m., n = 12 eyes/treatment.

Article Snippet: ERGs were obtained from animals 14 days post intravitreal injection of 100 μM SH-11037, 0.2 ng mouse anti-VEGF164 neutralizing antibody (R&D Systems, Minneapolis, MN, USA) + 0.3 μM SH-11037, or vehicle control and performed as previously described .

Techniques: Confocal Microscopy, Staining, Software

Journal: Scientific Reports

Article Title: A novel small molecule ameliorates ocular neovascularisation and synergises with anti-VEGF therapy

doi: 10.1038/srep25509

Figure Lengend Snippet: ( a ) The effects of 0.5 μM SH-11037, different concentrations of aflibercept, and SH-11037/aflibercept combinations on HREC proliferation were tested using an alamarBlue fluorescence assay, *** P < 0.001, one-way ANOVA with Tukey’s post hoc tests. Mean ± s.e.m., n = 3. Representative data from duplicate experiments. ( b ) Calculations of the excess % inhibition over highest single agent (HSA) activity and excess over Bliss additivity (see Methods). Values ≤ 0 are coloured in grey, values from 1–10 are in blue, values >10, indicating synergy, are yellow squares. ( c ) Representative images from confocal microscopy of agglutinin stained CNV lesions for combination treatment; 1 ng anti-VEGF/1 μM SH-11037 (Combo (1/1)) compared to individual SH-11037 and anti-VEGF164 treatments and vehicle control, scale bars = 50 μm. ( d ) Dose-dependent inhibition of the volume of CNV lesions by anti-VEGF164 injections. No difference was observed between anti-VEGF164 + vehicle compared to anti-VEGF164 alone ( e ) Quantification of CNV lesion volume shows a substitution of anti-VEGF164 by SH-11037. The combination of 1 ng anti-VEGF164 with 1 μM SH-11037 produces a similar effect to that observed by the 5 ng dose of anti-VEGF164 alone. The combination of 0.2 ng anti-VEGF164 with 0.3 μM SH-11037 (Combo (0.2/0.3)) is significantly different from individual treatments alone, and exceeds Bliss additivity and HSA by 36 and 37%, respectively, indicating synergy. Graphs in ( d , e ) are quantification of CNV lesion volumes from Z-stack images. * P <0.05, ** P <0.01, **** P <0.0001, ns; non-significant, one-way ANOVA, Tukey’s post hoc tests. Mean ± s.e.m., n = 12 eyes/treatment. ( f ) Representative mean ERG responses of combo (0.2/0.3) and vehicle treatments. ( g ) Quantification of scotopic a- and b- waves and photopic b-wave shows no difference in retinal function (stimulus: scotopic = 2.5, photopic = 25 cd∙s/m 2 ). P > 0.05, Student’s t-test. Mean ± s.e.m., n = 6 eyes/treatment.

Article Snippet: ERGs were obtained from animals 14 days post intravitreal injection of 100 μM SH-11037, 0.2 ng mouse anti-VEGF164 neutralizing antibody (R&D Systems, Minneapolis, MN, USA) + 0.3 μM SH-11037, or vehicle control and performed as previously described .

Techniques: Fluorescence, Inhibition, Activity Assay, Confocal Microscopy, Staining, Control

Journal: Molecular medicine reports

Article Title: Mechanical stress up-regulates RANKL expression via the VEGF autocrine pathway in osteoblastic MC3T3-E1 cells.

doi: 10.3892/mmr_00000088

Figure Lengend Snippet: Figure 1. The effect of mechanical stress on mRNA expression of VEGF-A, VEGFR-1, RANKL, OPG, M-CSF, ALP, collagen type I and MMP-13. (A) VEGF-A mRNA expression increased time-dependently from 6 h, and resulted in a 2.7-fold increase at 48 h. (B) VEGFR-1 mRNA expression increased slightly at 6, 12 and 24 h, and resulted in a 2.0-fold increase at 48 h. (C) RANKL mRNA expression increased time-dependently from 6 h, and resulted in a 3.6-fold increase at 48 h. (D) OPG mRNA expression increased at 24 and 48 h, and resulted in a 2.0-fold increase at 48 h. (E) M-CSF mRNA expression was unchanged at the 6, 12, 24 and 48 h time points. (F) ALP mRNA expression decreased time-dependently, and resulted in a 0.65-fold decrease at 48 h. (G) Collagen type I mRNA expression remained unchanged. (H) MMP-13 mRNA expression resulted in a 1.5-fold increase at 48 h only. Results are shown as the means ± SD (n=4). The control group at 6 h is defined as the standard (=1). Control, control group; MS, mechanical stress group.

Article Snippet: Anti-mouse VEGF neutralizing antibody (rmVEGF) (AF-493-NA; R&D Systems Inc.) was added to the culture medium 24 h before the application of the Flexercell Strain Unit.

Techniques: Expressing, Control

Journal: Molecular medicine reports

Article Title: Mechanical stress up-regulates RANKL expression via the VEGF autocrine pathway in osteoblastic MC3T3-E1 cells.

doi: 10.3892/mmr_00000088

Figure Lengend Snippet: Figure 2. Effect of mechanical stress on the protein concentration of VEGF and M-CSF. (A) VEGF protein concentration in the mechanical stress group increased time-dependently, showing significant differences compared with the control group at 6, 12, 24 and 48 h, respectively. (B) M-CSF protein concentration in the two groups increased time-dependently, and was lower in the mechanical stress group than in the control group at 12, 24 and 48 h. The means and standard deviations of the VEGF and M-CSF protein concentration are shown (n=4). Control, control group; MS, mechanical stress group.

Article Snippet: Anti-mouse VEGF neutralizing antibody (rmVEGF) (AF-493-NA; R&D Systems Inc.) was added to the culture medium 24 h before the application of the Flexercell Strain Unit.

Techniques: Protein Concentration, Control

Journal: Molecular medicine reports

Article Title: Mechanical stress up-regulates RANKL expression via the VEGF autocrine pathway in osteoblastic MC3T3-E1 cells.

doi: 10.3892/mmr_00000088

Figure Lengend Snippet: Figure 3. Neutralizing effect of anti-VEGF antibody on mRNA expression of VEGF-A, VEGFR-1, RANKL, OPG, M-CSF, ALP, collagen type I and MMP-13. VEGF-A mRNA expression was nearly equal to the control at 6 and 12 h and elevated at 24 and 48 h. (A) VEGF-A mRNA expression resulted in a 1.5-fold increase at 48 h. (B) VEGFR-1 mRNA expression was equivalent to the control at 6, 12, 24 and 48 h. (C) RANKL mRNA expression was nearly equal to the control at 6 h and elevated at 12, 24 and 48 h, and resulted in a 2.0-fold increase at 48 h. (D) OPG mRNA expression increased slightly at 24 and 48 h, and resulted in a 1.7-fold increase at 48 h. (E) M-CSF mRNA expression remained unchanged at 6, 12, 24 and 48 h. (F) ALP mRNA expression decreased time-dependently, and resulted in a 0.4-fold decrease at 48 h. (G and H) Collagen type I and MMP-13 mRNA expression was equivalent to control levels at 6, 12, 24 and 48 h. Results are shown as the means ± SD (n=4). The control group at 6 h is defined as the standard (=1). Control, control group; MS, mechanical stress group.

Article Snippet: Anti-mouse VEGF neutralizing antibody (rmVEGF) (AF-493-NA; R&D Systems Inc.) was added to the culture medium 24 h before the application of the Flexercell Strain Unit.

Techniques: Expressing, Control