Primary human brain vascular pericytes were cultured in Specialty Medium (ScienCell Research Laboratories, Carlsbad, CA) alone or supplemented with IFN-γ 30ng/mL (Peprotech, Rocky Hill, NJ) after 24 and 72 hours. After 96 hours, IL-6 and VEGF concentrations were analyzed in the culture supernatant by Luminex, and PDGFRβ (Biolegend, San Diego, CA) and cleaved caspase-3 (Cell Signaling, Danvers, MA) expression on pericytes were determined by flow cytometry.
Pdgfrβ
Manufactured by BioLegend
PDGFRβ is a cell surface receptor tyrosine kinase that binds to platelet-derived growth factor (PDGF). It plays a crucial role in cell proliferation, migration, and survival.
Automatically generated - may contain errors
Lab products found in correlation
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Ifn γ, by Thermo Fisher Scientific (1 mentions)
Specialty medium, by ScienCell (1 mentions)
Lsrii sorp, by BD (1 mentions)
Sca 1, by Thermo Fisher Scientific (1 mentions)
Pdgfrα, by BioLegend (1 mentions)
Facscanto 2 flow cytometer, by BD (1 mentions)
Facsdiva software, by BD (1 mentions)
Cd105, by Thermo Fisher Scientific (1 mentions)
3 protocols using pdgfrβ
1
Pericyte Vascular Inflammatory Response
2
Immunophenotyping of Isolated Cells
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Isolated cells were incubated with primary antibodies for 40 min at 4 °C in phosphate-buffered saline (PBS) supplemented with 2% FBS and 2 mM ethylenediaminetetraacetic acid (EDTA). We used the following directly conjugated antibodies: CD29 (R&D Systems), CD44 (BioLegend), CD90.2 (BioLegend), CD31 (BioLegend), CD34 (BioLegend), stem cell antigen 1 (Sca-1; e-Bioscience), PDGFRα (BioLegend), and PDGFRβ (BioLegend). All stainings were controlled with appropriate isotope control antibodies. Analysis was performed on a SORP LSRII (Becton Dickinson) equipped with five lasers and data were collected with FACS DIVA software. Analysis was performed using FlowJo™ 10.0.8 (Treestar, Ashland, OR). All measurements were performed with three biological replicates.
3
Phenotypic Characterization of Bone Marrow-Derived Mesenchymal Stem Cells
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APCs were stained for surface antigen expression using combinations of the following antibodies to confirm typical phenotype: CD105 (Life Technologies), CD90, CD73, PDGFRβ (Biolegend), CD44 (eBioscience), CD31 (BD Biosciences), CD34, and CD45 (Miltenyi)5 (link), 10 (link). Analysis was performed using a FACS Canto II flow cytometer and FACS Diva software (both BD Biosciences). BM-MSCs antigenic characterization was also routinely performed by flow cytometry. Primers and antibodies are listed in Supplementary Tables VI and VII , respectively.
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