Mouse PDGF R beta Antibody Best Seller

Detection of Mouse PDGF R beta by Immunocytochemistry/Immunofluorescence

VCAM1 is involved in the regulation of proliferation and morphological changes in oligodendrocytes.(a) Tissue lysates from 7-day-old NG2-Cre-driven VCAM1 conditional knockout (VCAM1fl/fl; Ng2-Cre) or control (Ctrl) mouse spinal cords or 11-day-old whole brains were immunoblotted with an antibody against MBP, CNPase, VCAM1 or actin. Data are representative of three experiments. (b–d) Antibodies against Ki67 (green) and Olig2 (red) were used for co-staining in 2-day-old spinal cord cross sections. Data are representative. The scale bars indicate 200 μm. The number of Olig2+ cells per one square millimetre was counted. Data were evaluated using Student’s t-test (**P=0.000528; n=19 slices of two independent experiments). The percentage of Ki67+ cells among the Olig2+ cells is shown. Data were evaluated using Student’s t-test (**P=0.00807; n=6 slices of two independent experiments). (e) Antibodies against Ki67 and NG2 were used for co-staining in 2-day-old spinal cord. The percentage of Ki67+ cells among the NG2+ cells per one square millimetre is shown. Data were evaluated using Student’s t-test (*P=0.0275; n=6 slices of two independent experiments). (f–h) Knock-down efficiencies of shRNAs were confirmed by immunoblotting. Data are representative of three experiments. Oligodendrocytes were transfected with VCAM1 shRNA or control, co-cultured with neurons for 2 days, and co-stained with antibodies against NF (green) and NG2 (red). Magnified images of the dotted boxes (i, attached; ii, aligned) are shown below. Data are representative. Scale bar, 100 μm. The percentage of NG2+ cells whose process tips or cell bodies were attached to axons or aligned along axons is shown. Data were evaluated using Student’s t-test (**P=0.00109 (attached cells) or P=0.00982 (aligned cells); n=6 areas of three experiments). (i,k) Oligodendrocytes were cultured on dishes coated with or without recombinant alpha4 beta1 integrin in a growth medium containing PDGF and bFGF. Cells were co-stained with an anti-MBP antibody (red) and DAPI (blue) (a) or an anti-PDGFR alpha antibody (green) (b). Data are representative. Scale bar, 100 μm. The percentage of MBP+ cells is shown. Data were evaluated using Student’s t-test (**P=1.14E−10; n=6 areas of two independent experiments). (j,l) Oligodendrocytes transfected with VCAM1#1 or control shRNA were cultured on recombinant alpha4 beta1 integrin-coated dishes in the differentiation medium and co-stained with an anti-MBP antibody (red) and DAPI (blue). Data are representative. Scale bar, 100 μm. The percentage of MBP+ cells is shown. Data were evaluated using Student’s t-test (**P=1.82E−21; n=6 areas of two independent experiments). (m–o) Antibodies against Ki67 (green) and O4 (red) were used for co-staining in 7-day-old spinal cord. DAPI staining is also shown. Data are representative. Scale bar, 100 μm. The number of O4+ cells was counted. Data were evaluated using Student’s t-test (**P=5.87E−05 (P7) or P=5.87E−06 (P14); n=12 slices of two independent experiments). The percentage of Ki67− or Ki67+ cells among the O4+ cells is shown. Data were evaluated using Student’s t-test (**P=0.00344 (P7, Ki67−), 0.00344 (P7, Ki67+) or 0.000580 (P14, Ki67+) *P=0.0494 (P14, Ki67−); n=12 slices of two independent experiments). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27876794), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse PDGF R beta 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 PDGF R beta in Mouse Embryo Brain.

Formalin-fixed paraffin-embedded tissue sections of mouse embryo were probed for PDGFRb mRNA (ACD RNAScope Probe, catalog #411388; Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using goat anti-mouse PDGFRb polyclonal antibody (R&D Systems catalog # AF1042) at 1.7ug/mL with overnight incubation at 4 degrees Celsius 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 developing vasculature.