Phycoerythrin cy7 anti mouse cd31 antibody

CD31^hiEMCN^hi cells in the femur and tibia were measured by flow cytometry as previously described (38 (link)). Briefly, we dissected the bones, made small cuts at both sides, and placed them in the sectioned tips within the centrifuge tubes. Then, the tubes were centrifuged at 10,000g for 15 s to get whole bone marrow cells. After ACK lysis, cells were incubated with phycoerythrin/Cy7 anti-mouse CD31 antibody (BioLegend), EMCN monoclonal antibody (eBioscience), fluorescein isothiocyanate (FITC) anti-mouse TER-119/erythroid cell antibody (BioLegend), and FITC anti-mouse CD45 antibody (BioLegend). Cell sorting was performed with CyAn ADP Analyzer (Beckman Coulter), and CD31⁺CD45⁻Ter119⁻ cells were considered bone marrow endothelial cells.

CD31^hiEMCN^hi cells in the bone were detected by flow cytometry. The femora were first removed from the mice without the epiphysis region. Then, the muscles and periosteum were removed from the bone. The samples were crushed and digested to obtain suspensions of bone marrow cells. The cells were incubated with phycoerythrin/Cy7 anti-mouse CD31 antibody (102417, BioLegend), Emcn monoclonal antibody [eBioV.7C7 (V.7C7), eBioscience], eFluor 660 viability dye, eBioscience (50585182, Invitrogen), fluorescein isothiocyanate (FITC) anti-mouse TER-119/erythroid cells antibody (116205, BioLegend), and FITC anti-mouse CD45 antibody (103107, BioLegend). A fluorescence-activated cell sorter (FACSAria, BD Biosciences) was used for analyses, and CD31⁺CD45⁻Ter119⁻ cells were considered bone marrow endothelial cells.