polyclonal antibodies against vegfr2  (Cell Signaling Technology Inc)

Journal: Frontiers in Pharmacology

Article Title: Dihydroartemisinin Inhibits Laser-Induced Choroidal Neovascularization in a Mouse Model of Neovascular AMD

doi: 10.3389/fphar.2022.838263

Figure Lengend Snippet: VEGFR2 and NF-κB p65 expression at the site of choroidal neovascularization (CNV) lesions analyzed by immunostaining images. The CNV lesion was indicated by the red ellipse. (A) The sections were fluorescently labeled with VEGFR2 antibody (green) and isolectin B4 (red). VEGFR2 had a little positive staining in retinal capillary endothelial cells in both the vehicle and dihydroartemisinin (DHA) groups. However, in CNV formation spots, the positive staining area of VEGFR2 in the DHA group was smaller than that in the vehicle group (white arrow). (B) The sections were fluorescently labeled with NF-κB p65 antibody (green) and isolectin B4 (red). The positive staining area of NF-κB p65 in the DHA group was significantly reduced compared to that in the vehicle group, which was mainly seen in CNV areas (yellow arrow). GCL, Ganglion Cell Layer; INL, Inner Nuclear Layer; ONL, Outer Nuclear Layer. Scale bar, 50 μm.

Article Snippet: The sections were stained with monoclonal antibodies against NF-κB p65 (CST, 8,242, Beverly, MA, United States) or with polyclonal antibodies against VEGFR2 (CST, 2,478, Beverly, MA, United States) at 4 °C overnight and then incubated with corresponding secondary antibodies conjugated to Alexa Fluor 488 (Invitrogen, United States) for 1 h at room temperature in the dark, while the nuclei were stained with 4′6-diamidino-2-phenylindole (DAPI; Sigma Aldrich).

Techniques: Expressing, Immunostaining, Labeling, Staining

Journal: Frontiers in Pharmacology

Article Title: Dihydroartemisinin Inhibits Laser-Induced Choroidal Neovascularization in a Mouse Model of Neovascular AMD

doi: 10.3389/fphar.2022.838263

Figure Lengend Snippet: The impact of DHA on the protein expression levels of the NF-κB family, VEGF, and VEGFR2 in the RPE-choroid complex of a laser-induced CNV mouse model. Mice were intragastrically administered vehicle or DHA once a day for 12 days. (A) The typical Western blot results showed changes in NF-κB family, VEGF, and VEGFR2 protein expression. (B) The quantitative analysis results demonstrated that the expression levels of NF-κB p65, NF-κB1 p50, Rel B, VEGF, and VEGFR2 were markedly increased in the vehicle group compared to the normal control group. DHA significantly inhibited the expression of NF-κB p65, NF-κB1 p50, VEGF, and VEGFR2. n = 3 experiments; * p < .05 vs Nor; # p < .05 vs Laser + Vehicle; one-way ANOVA followed by Tukey’s test. CNV: Choroidal Neovascularization; DHA: Dihydroartemisinin.

Article Snippet: The sections were stained with monoclonal antibodies against NF-κB p65 (CST, 8,242, Beverly, MA, United States) or with polyclonal antibodies against VEGFR2 (CST, 2,478, Beverly, MA, United States) at 4 °C overnight and then incubated with corresponding secondary antibodies conjugated to Alexa Fluor 488 (Invitrogen, United States) for 1 h at room temperature in the dark, while the nuclei were stained with 4′6-diamidino-2-phenylindole (DAPI; Sigma Aldrich).

Techniques: Expressing, Western Blot, Control

Journal: Frontiers in Pharmacology

Article Title: Dihydroartemisinin Inhibits Laser-Induced Choroidal Neovascularization in a Mouse Model of Neovascular AMD

doi: 10.3389/fphar.2022.838263

Figure Lengend Snippet: Schematic illustration of the mechanisms of the anti-CNV effects of DHA. In addition to the downregulation of VEGF expression, DHA inhibits VEGFR2 expression and angiogenesis through suppression of the NF-κB pathway.

Article Snippet: The sections were stained with monoclonal antibodies against NF-κB p65 (CST, 8,242, Beverly, MA, United States) or with polyclonal antibodies against VEGFR2 (CST, 2,478, Beverly, MA, United States) at 4 °C overnight and then incubated with corresponding secondary antibodies conjugated to Alexa Fluor 488 (Invitrogen, United States) for 1 h at room temperature in the dark, while the nuclei were stained with 4′6-diamidino-2-phenylindole (DAPI; Sigma Aldrich).

Techniques: Expressing

Journal: Cell Cycle

Article Title: Cancer cell stemness, responses to experimental genotoxic treatments, cytomegalovirus protein expression and DNA replication stress in pediatric medulloblastomas

doi: 10.1080/15384101.2020.1728025

Figure Lengend Snippet: Immunohistochemical data for marker expression in human MB*.

Article Snippet: The primary antibodies used for this immunohistochemical analysis included the following reagents: mouse monoclonal antibody against CD133 (AC133 pure, Miltenyi Biotec, diluted 1:50), mouse monoclonal antibody to CD15 (NCL-CD15, Leica, diluted 1:100), rabbit polyclonal antibody against VEGFR2 (Cell Signaling, diluted 1:250), and rabbit polyclonal antibody to phosphorylated RPA32 (Ser 4/8, NBP1-23017, Novus Biologicals, 1:2000).

Techniques: Immunohistochemical staining, Marker, Expressing

Journal: Scientific Reports

Article Title: Vascular Endothelial Growth Factor in Cartilage Development and Osteoarthritis

doi: 10.1038/s41598-017-13417-w

Figure Lengend Snippet: Effects of VEGF on OA progression. DMM or sham surgery was performed in 24 week-old wild type C57BL/6J mice and the outcome analyzed 8 weeks later. The mice received intra-articular anti-VEGF antibody or phosphate buffered saline (PBS) at weeks 1, 2, 3 and 4 post DMM ( A ) ( n = 10). Sections of knee joints were stained with Safranin O-fast green, high magnification insets show articular cartilage details ( B ). Assessment of cartilage degradation by OARSI grading scores ( n = 6) ( C ). Immunofluorescence staining of phosphorylated VEGFR2 (pVEGFR2) close to the articular surface; high magnification inset ( D ), in synovium ( F ) and dorsal root ganglia (DRG) ( I ). Immunofluorescence staining for phosphorylated VEGFR1 (pVEGFR1) in the DRG ( H ); numbers of pVEGFR2 positive cells in articular surface region ( E ) and synovium ( G ) are reduced in anti-VEGF treated joints. Numbers of pVEGFR1 positive cells in DRG neurons are significantly reduced in anti-VEGF treated joints ( J ); reduction of pVEGFR2 positive cells is not significant ( K ). Red fluorescence represents pVEGFR2 ( D,F and I ) or pVEGFR1 ( H ); green fluorescence represents neuronal nuclear antigen (NeuN) ( H and I ); blue represents DAPI nuclear staining. Scale bars, 250 μm; *P < 0.05. Data represent mean ± SEM. Unpaired Student’s t-test was used.

Article Snippet: After antigen retrieval, the sections were incubated overnight with rabbit polyclonal antibody against pVEGFR2 (orb99143, Biorbyt) at 1:100 dilution.

Techniques: Saline, Staining, Immunofluorescence, Fluorescence