Mouse VEGFR2/KDR/Flk-1 Antibody

Detection of Mouse VEGFR2/KDR/Flk-1 by Immunocytochemistry/Immunofluorescence

BM-PC express Notch pathway ligands/receptors and show increased expression of notch downstream targets during endothelial differentiation.A. Expression of Notch receptors and ligands in BM-PC was detected by RT-PCR. B. Expression of Notch downstream targets (Hes 1, Hey 1 and 2) was detected at different time points during BM-PC endothelial differentiation by quantitative real-time PCR. C. Representative images (×200) of BM-PC at day 20 of culture showing positivity for endothelial lineage specific markers, acetylated LDL, CD31 , Flk-1 and vWF with DAPI nuclear counterstaining in blue. D. Quantification of BM-PC positive cells for acetylated LDL, CD31 , Flk-1 and VWF after 20 days of culture. Each experiment was performed in triplicate and the mean presented (n = 3). Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0003752), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse VEGFR2/KDR/Flk-1 by Immunohistochemistry

Vascular alterations after intraocular VEGF-A injection. a Morphology of IB4-stained P6 wild-type retinal vessels at 4 h after administration of human VEGF-A165 (0.5 µl at a concentration of 5 μg μl−1). Note blunt appearance of the vessel front after VEGF-A injection but not for vehicle (PBS) control. Scale bar, 200 µm. b Quantitation of sprouts and filopodia at the front of the P6 vessel plexus after injection of VEGF-A165 or vehicle control. Error bars, s.e.m. p-values, Student’s t-test. c PDGFR beta+ (green) pericytes are unaffected by short-term VEGF-A administration, whereas VEGFR2 immunosignals (white) are increased in IB4+ (red) ECs (arrowheads). Images shown correspond to insets in a. Scale bar, 100 µm. d Quantitation of VEGFR2 immunosignals intensity in the peripheral plexus of P6 retinas after injection of VEGF-A165 or vehicle control. Error bars, s.e.m. p-values, Student’s t-test. e Confocal images showing increased Esm1 immunostaining (white) in IB4+ (red) ECs in the peripheral plexus (arrowheads) after VEGF-A injection in P6 pups. Scale bar, 200 µm. f VEGF-A165 injection-mediated increase of Esm1 immunosignals (normalized to IB4+ EC area) in the peripheral capillary plexus but not at the edge of the angiogenic front in comparison to PBS-injected controls at P6. Error bars, s.e.m. p-values, Student’s t-test. NS, not statistically significant. g Short-term VEGF-A165 administration leads to clustering of Erg1+ (green) and IB4+ (red) ECs, as indicated, in thick sprout-like structures of P6 retinas. Panels in the center and on the right (scale bar, 20 µm) show higher magnification of the insets on the left (scale bar, 100 µm). Dashed lines in panels on the right outline IB4+ vessels. h Quantitation of EC density in the leading front vessel and emerging sprouts of the P6 angiogenic front after injection of VEGF-A165 or vehicle control. Error bars, s.e.m. p-values, Student’s t-test Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29146905), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse VEGFR2/KDR/Flk-1 by Immunocytochemistry/Immunofluorescence

Inactivation of Flt1 in PDGFR beta+ cells. a Experimental scheme of tamoxifen administration for the generation of Flt1iPC mutants. b P6 control, Flt1iPC/+ and Flt1iPC retinas stained with isolectin B4 (IB4). Dashed circles indicate vessel-covered (yellow) and peripheral avascular (white) areas in the overview pictures (top). Scale bar, 500 µm. c Quantitation of body weight and radial outgrowth of the retinal vasculature in control, Flt1iPC/+ and Flt1iPC P6 pups. Error bars, s.e.m. p-values, one-way ANOVA. NS, not statistically significant. d Confocal images of the IB4-stained P6 control, Flt1iPC/+ and Flt1iPC retinal angiogenic front illustrating differences in sprout number and morphology. Scale bar, 100 µm. e Quantitation of sprouts and filopodia in P6 control, Flt1iPC/+ and Flt1iPC retinas. Error bars, s.e.m. p-values, one-way ANOVA and Tukey’s multiple comparison test. NS, not statistically significant. f Confocal images of IB4 (red), Erg1 (green) and VEGFR2 (white) stained P6 retinas highlighting the accumulation of EC nuclei and enhanced VEGFR2 immunosignals (arrowheads) in Flt1iPC sprouts. Vessels are outlined by dashed lines on the right panel. Scale bar, 100 µm. g Quantitation of EC proliferation (EdU+ Erg1+) at the angiogenic front, EC density in sprouts and leading front vessel and VEGFR2 immunosignals intensity in the angiogenic front of control and Flt1iPC P6 retinas. Error bars, s.e.m. p-values, Student’s t-test. h Esm1 (white) expression (arrowheads) in the angiogenic front (IB4+, red, first two columns) and detection of desmin+ pericytes (green, third column) in P6 control and Flt1iPC retinas. Scale bar, 100 µm. i Quantitation of Esm1+ proportion relative to vascular area (IB4+) in the angiogenic front of control and Flt1iPC P6 retinas. Error bars, s.e.m. p-values, Student’s t-test. j Confocal images of P6 retinas stained for NG2 (green) and IB4 (red) showing no significant changes in pericyte coverage in the front (first two columns) or the remodeling plexus around veins (v) or arteries (a) (last two columns). Scale bar, 100 µm. k, l Quantitation of pericyte coverage k and relative gene expression by qPCR on whole lysates l in control and Flt1iPC P6 retinas. Error bars, s.e.m. p-values, Student’s t-test. NS, not statistically significant Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29146905), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of VEGFR2/KDR/Flk-1 in Mouse Kidney.

Formalin-fixed paraffin-embedded tissue sections of mouse kidney were probed for VEGFR2 mRNA (ACD RNAScope Probe, catalog #414818, Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using goat anti-mouse VEGFR2 polyclonal antibody (R&D Systems catalog # AF644) at 1.7ug/mL with 1 hour incubation at room temperature followed by incubation with anti-goat IgG VisUCyte HRP Polymer Antibody (Catalog # VC004) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to glomeruli and fibroblasts.

Detection of Mouse Mouse VEGFR2/KDR/Flk-1 Antibody by Immunocytochemistry/ Immunofluorescence

BM-PC express Notch pathway ligands/receptors and show increased expression of notch downstream targets during endothelial differentiation.A. Expression of Notch receptors and ligands in BM-PC was detected by RT-PCR. B. Expression of Notch downstream targets (Hes 1, Hey 1 and 2) was detected at different time points during BM-PC endothelial differentiation by quantitative real-time PCR. C. Representative images (×200) of BM-PC at day 20 of culture showing positivity for endothelial lineage specific markers, acetylated LDL, CD31 , Flk-1 and vWF with DAPI nuclear counterstaining in blue. D. Quantification of BM-PC positive cells for acetylated LDL, CD31 , Flk-1 and VWF after 20 days of culture. Each experiment was performed in triplicate and the mean presented (n = 3). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/19015735), licensed under a CC-BY license. Not internally tested by R&D Systems.