Journal: Investigative ophthalmology & visual science

Article Title: Apatinib, an Inhibitor of Vascular Endothelial Growth Factor Receptor 2, Suppresses Pathologic Ocular Neovascularization in Mice.

doi: 10.1167/iovs.17-21416

Figure Lengend Snippet: FIGURE 1. Apatinib inhibits VEGF-induced phosphorylation of VEGFR2 and ERK in HRMECs. (A) Representative Western blotting images and (B, C) densitometric analysis of phosphorylated VEGFR2 (p-VEGFR2; B) and phosphorylated ERK (p-ERK; C) are illustrated. HRMECs treated with apatinib (1 lM) and untreated cells were stimulated with rhVEGF (100 ng/mL) or PBS for 20 minutes. Cell lysates were subjected to Western blotting using primary IgGs against p-VEGFR2 and p-ERK. The membranes were then stripped and reprobed with IgGs against VEGFR2, ERK, and b-actin. Band intensities of p-VEGFR2 and p-ERK were normalized to the band intensities of VEGFR2 and ERK, respectively (*P < 0.05 versus PBS, #P < 0.05 versus VEGF only, n ¼ 3).

Article Snippet: After quenching endogenous peroxidase activity and blocking with 10% normal goat serum, tissue sections were incubated with primary IgGs against p-VEGFR2 (Origene) or glial fibrillary acidic protein (GFAP; Dako), and then incubated with the appropriate fluorescent secondary IgG.

Techniques: Phospho-proteomics, Western Blot

Journal: Investigative ophthalmology & visual science

Article Title: Apatinib, an Inhibitor of Vascular Endothelial Growth Factor Receptor 2, Suppresses Pathologic Ocular Neovascularization in Mice.

doi: 10.1167/iovs.17-21416

Figure Lengend Snippet: FIGURE 3. Intravitreal injection of apatinib decreases retinal neovascularization in mice with OIR. (A) Schematic diagram of the OIR experiment. On P12, mice received a single intravitreal injection of apatinib (1 lg in 1 lL DMSO) or DMSO (1 lL; contralateral control). Five days later (on P17), the eyes were harvested for further analysis. (B) Representative images of whole-mounted retinas with neovascular tufts highlighted in white, and quantification of retinal neovascularization. Retinal vasculature was visualized by staining with isolectin B4 (red). The extent of retinal neovascularization was calculated by dividing the number of pixels in the neovascular tuft area by the number of pixels in the total retinal area. All data are presented as the mean 6 SEM (**P < 0.01, n ¼ 4 mice per group). Scale bar: 500 lm. (C) Immunofluorescence staining was performed using anti-p-VEGFR2 IgGs (green) and isolectin B4 (red) on cryosections prepared from the eyecups of mice with OIR. The arrowheads indicate the double labeling of p-VEGFR2 and isolectin B4. Sections stained with irrelevant nonspecific IgGs are included as negative controls (NC). The nuclei are shown in blue (DAPI). Representative images were selected from three independent experiments with similar results. Scale bar: 50 lm.

Article Snippet: After quenching endogenous peroxidase activity and blocking with 10% normal goat serum, tissue sections were incubated with primary IgGs against p-VEGFR2 (Origene) or glial fibrillary acidic protein (GFAP; Dako), and then incubated with the appropriate fluorescent secondary IgG.

Techniques: Injection, Control, Staining, Labeling

Journal: Investigative ophthalmology & visual science

Article Title: Apatinib, an Inhibitor of Vascular Endothelial Growth Factor Receptor 2, Suppresses Pathologic Ocular Neovascularization in Mice.

doi: 10.1167/iovs.17-21416

Figure Lengend Snippet: FIGURE 4. Intravitreal injection of apatinib decreases CNV formation in mice with laser-induced CNV. (A) Schematic diagram of the laser-induced CNV experiment. Immediately after laser photocoagulation, mice received a single intravitreal injection of apatinib (1 lg in 1 lL DMSO) or DMSO (1 lL; contralateral control). Two weeks later, the eyes were harvested for further analysis. (B) Representative images of flat-mounted choroids with CNV and quantification of the isolectin B4–positive CNV area. The posterior eyecups composed of RPE, choroid, and sclera were flat-mounted and stained with isolectin B4 (red). The CNV areas were quantified by measuring the fluorescence intensity of images with isolectin B4–positive areas. All data are presented as the mean 6 SEM (**P < 0.01, n¼4 mice per group). (C) Immunofluorescence staining was performed using anti-p-VEGFR2 IgGs (green) and isolectin B4 (red) on cryosections prepared from the eyecups of mice with CNV. The arrowheads indicate the double labeling of p- VEGFR2 and isolectin B4 in CNV lesions (white dashed circles). Sections stained with irrelevant nonspecific IgGs are included as negative controls (NC). The nuclei are shown in blue (DAPI). Representative images were selected from three independent experiments with similar results. Scale bars: 100 lm.

Article Snippet: After quenching endogenous peroxidase activity and blocking with 10% normal goat serum, tissue sections were incubated with primary IgGs against p-VEGFR2 (Origene) or glial fibrillary acidic protein (GFAP; Dako), and then incubated with the appropriate fluorescent secondary IgG.

Techniques: Injection, Control, Staining, Labeling

Journal: Cell reports

Article Title: Human Pluripotent Stem Cell-Derived Multipotent Vascular Progenitors of the Mesothelium Lineage Have Utility in Tissue Engineering and Repair

doi: 10.1016/j.celrep.2019.02.016

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit Polyclonal anti-Flk1-1 , Acris Antibodies GmbH , Cat# AP02618PU-S; RRID: AB_1624459.

Techniques: Control, Recombinant, Clinical Proteomics, Modification, Membrane, Sterility, Reverse Transcription, DNA Methylation Assay, Flow Cytometry, Methylation, Software, Imaging, Cell Culture

Journal: Journal of the Chinese Medical Association : JCMA

Article Title: The effects of pioglitazone in cirrhotic rats with hepatopulmonary syndrome.

doi: 10.1016/j.jcma.2017.08.010

Figure Lengend Snippet: Fig. 5. Pulmonary protein expressions of VEGF, VEGFR2, p-VEGFR2, p-VEGFR2/VEGFR2, eNOS, p-eNOS, p-eNOS/eNOS, iNOS in vehicle- and pioglitazone- treated rats with CBDL-induced HPS. Pioglitazone significantly down-regulated VEGF expression and p-eNOS/eNOS as compared with vehicle ( p < 0.05).

Article Snippet: The protein extracts were incubated with the primary antibody [anti-VEGF rabbit polyclonal antibody (Gene Tex, Irvine, CA, USA); anti-VEGFR-2 (1:500; Millipore Corporation, Billerica, MA, USA); anti-p-VEGFR2 rabbit polyclonal antibody (OriGene, Rockville, USA); anti-eNOS, -iNOS (1:1000; Millipore Corporation, Billerica, MA, USA); anti-peNOS (1:1000; Cell Signaling Technology, Danvers, MA, USA)].

Techniques: Expressing

Journal: Molecular Medicine Reports

Article Title: Effect of Melilotus suaveolens extract on pulmonary microvascular permeability by downregulating vascular endothelial growth factor expression in rats with sepsis

doi: 10.3892/mmr.2015.3146

Figure Lengend Snippet: M. suaveolens extract blocked the expression of VEGF and NF-κB mRNA in lung tissue. Groups of mice were challenged with cecal ligation and puncture and treated with M. suaveolens extract 24 h later. The left lung tissues were homogenized and total RNA was extracted using TRIzol reagent and assayed by quantitative polymerase chain reaction. (A) Representative gels assessing (a) VEGF and (b) NF-κB levels are demonstrated. Lane 1, marker; lane 2, normal control group; lane 3, sham operation group; lane 4, (untreated) sepsis group and lane 5, treatment group. (B) Statistical summary of the densitometric analysis of VEGF and NF-κB mRNA expression in rats from the four groups; (a) VEGF mRNA and (b) NF-κB mRNA. Data are represented as the mean ± standard deviation of one experiment consisting of three replicates. ** P<0.01 vs. sham operation and normal control groups; ## P<0.01 vs. (untreated) sepsis group. VEGF, vascular endothelial growth factor; NF-κB, nuclear factor kappa B.

Article Snippet: Primary antibodies used included rabbit anti-VEGF monoclonal antibody (1:400), rabbit anti-NF-κB65 monoclonal antibody (1:400) (Boster Biological Technology, Ltd) and mouse anti- GAPDH monoclonal antibody (1:400) (Santa Cruz Biotechnology, Inc).

Techniques: Expressing, Ligation, Real-time Polymerase Chain Reaction, Marker, Control, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Effect of Melilotus suaveolens extract on pulmonary microvascular permeability by downregulating vascular endothelial growth factor expression in rats with sepsis

doi: 10.3892/mmr.2015.3146

Figure Lengend Snippet: Effect of M. suaveolens extract on the expression of VEGF and NF-κB65 protein in lung tissue. Groups of mice were challenged with cecal ligation and puncture and treated with M. suaveolens extract 24 h later. The expression of VEGF, NF-κβ65 and GAPDH was detected by western blotting using specific antibodies. GAPDH protein was used an internal control. (A) Representative western blot analysis demonstrated the levels of VEGF and NF-κB65 protein expression in rats from the four groups; (a) normal control group; (b) sham operation group; (c) (untreated) sepsis group and (d) treatment group. (B) Quantification of the blots by densitometric analysis of (a) VEGF and (b) NF-κB65 protein expression in rats from the four groups. Data are presented as the mean ± standard deviation of one experiment consisting of three replicates. ** P<0.01 vs. the sham operation group and normal control group; # P<0.05 vs . (untreated) sepsis group. VEGF, vascular endothelial growth factor; NF-κB, nuclear factor kappa B.

Article Snippet: Primary antibodies used included rabbit anti-VEGF monoclonal antibody (1:400), rabbit anti-NF-κB65 monoclonal antibody (1:400) (Boster Biological Technology, Ltd) and mouse anti- GAPDH monoclonal antibody (1:400) (Santa Cruz Biotechnology, Inc).

Techniques: Expressing, Ligation, Western Blot, Control, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Effect of Melilotus suaveolens extract on pulmonary microvascular permeability by downregulating vascular endothelial growth factor expression in rats with sepsis

doi: 10.3892/mmr.2015.3146

Figure Lengend Snippet: Effect of M. suaveolens extract on the protein expression of VEGF and NF-κB65 in rat lungs 24 h following cecal ligation and puncture-induced acute lung injury. Immunostaining was performed on lung sections following antigen retrieval using Retrievagen. (A) Representative immunostaining revealed VEGF and NF-κβ65-positive expression in rats from the four groups: (a–c) Expression of positive VEGF in the (a) sham operation group; (b) control group; (c) treatment group); (d–f) Expression of positive NF-κB65 in the (d) sham operation group; (e) (untreated) sepsis group; (f) treatment group (magnification, ×200). (B) Quantification of the images by densitometric analysis of (a) VEGF and (b) NF-κB-positive protein expression in rats from four groups. All values are expressed as the mean ± standard deviation. ** P<0.01 vs. the sham operation group; ## P<0.01 vs. (untreated) sepsis group. VEGF, vascular endothelial growth factor; NF-κB, nuclear factor kappa B.

Article Snippet: Primary antibodies used included rabbit anti-VEGF monoclonal antibody (1:400), rabbit anti-NF-κB65 monoclonal antibody (1:400) (Boster Biological Technology, Ltd) and mouse anti- GAPDH monoclonal antibody (1:400) (Santa Cruz Biotechnology, Inc).

Techniques: Expressing, Ligation, Immunostaining, Control, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Effect of Melilotus suaveolens extract on pulmonary microvascular permeability by downregulating vascular endothelial growth factor expression in rats with sepsis

doi: 10.3892/mmr.2015.3146

Figure Lengend Snippet: Effect of M. suaveolens extract on plasma levels of VEGF, TNF-α, IL-6, IL-1β, IL-4 and IL-10 levels in plasma. Groups of mice were challenged with cecal ligation and puncture, and treated with M. suaveolens extract 24 h later. (A) VEGF, (B) TNF-α, (C) IL-6, (D) IL-1β, (E) IL-10 and (F) IL-4 levels in plasma were determined by ELISA. Data are presented as the mean ± standard deviation of one experiment consisting of three replicates. * P<0.05, ** P<0.01 vs. the sham operation group and normal control group; # P<0.05, ## P<0.01 vs. (untreated) sepsis group. VEGF, vascular endothelial growth factor; NF-κB, nuclear factor kappa B; TNF-α, tumor necrosis factor-α; IL, interleukin.

Article Snippet: Primary antibodies used included rabbit anti-VEGF monoclonal antibody (1:400), rabbit anti-NF-κB65 monoclonal antibody (1:400) (Boster Biological Technology, Ltd) and mouse anti- GAPDH monoclonal antibody (1:400) (Santa Cruz Biotechnology, Inc).

Techniques: Clinical Proteomics, Ligation, Enzyme-linked Immunosorbent Assay, Standard Deviation, Control

Journal: Molecular Medicine Reports

Article Title: Effect of Melilotus suaveolens extract on pulmonary microvascular permeability by downregulating vascular endothelial growth factor expression in rats with sepsis

doi: 10.3892/mmr.2015.3146

Figure Lengend Snippet: Administration of M. suaveolens extract attenuated lipopolysaccharide-induced pulmonary inflammation. Groups of mice were challenged with cecal ligation and puncture, and treated with M. suaveolens extract 24 h later. (A) VEGF, (B) TNF-α, (C) IL-6, (D) IL-1β, (E) IL-10 and (F) IL-4 levels in bronchoalveolar lavage were determined by ELISA. Data are presented as the mean ± standard deviation of one experiment consisting of three replicates. ** P<0.01 vs. the sham operation group and normal control group; # P<0.05, ## P<0.01 vs. (untreated) sepsis group. VEGF, vascular endothelial growth factor; NF-κB, nuclear factor kappa B; TNF-α, tumor necrosis factor-α; IL, interleukin.

Article Snippet: Primary antibodies used included rabbit anti-VEGF monoclonal antibody (1:400), rabbit anti-NF-κB65 monoclonal antibody (1:400) (Boster Biological Technology, Ltd) and mouse anti- GAPDH monoclonal antibody (1:400) (Santa Cruz Biotechnology, Inc).

Techniques: Ligation, Enzyme-linked Immunosorbent Assay, Standard Deviation, Control

Journal: Molecular Medicine Reports

Article Title: Effect of Melilotus suaveolens extract on pulmonary microvascular permeability by downregulating vascular endothelial growth factor expression in rats with sepsis

doi: 10.3892/mmr.2015.3146

Figure Lengend Snippet: Administration of M. suaveolens extract reduces CLP-induced lung permeability. Rats were treated as indicated and (A) FITC-labeled albumin in the bronchoalveolar lavage fluid, (B) water content of lung tissue and (C) W/D lung weight ratio were determined 24 h following CLP challenge. Data are presented as the mean ± standard deviation of one experiment consisting of three replicates. * P<0.05, ** P<0.01 vs. the sham operation and normal control groups; ## P<0.01 vs. (untreated) sepsis group. VEGF, vascular endothelial growth factor; NF-κB, nuclear factor kappa B; TNF-α, tumor necrosis factor-α; IL, interleukin; CLP, cecal ligation and puncture; W/D, wet/dry; FITC, fluorescein isothiocyanate.

Article Snippet: Primary antibodies used included rabbit anti-VEGF monoclonal antibody (1:400), rabbit anti-NF-κB65 monoclonal antibody (1:400) (Boster Biological Technology, Ltd) and mouse anti- GAPDH monoclonal antibody (1:400) (Santa Cruz Biotechnology, Inc).

Techniques: Permeability, Labeling, Standard Deviation, Control, Ligation