Cd45 pe cytm7

Single cell suspension dissociated from S1M allografts and primary mouse cells were stained with Sca‐1 FITC (553335; BD Pharmingen), CD133 PE (12‐1331‐82; eBioScience), CD44 FITC (11‐0441‐81; eBioScience), CD45 PE‐CyTM7 (552848; BD Pharmingen), rat IgG isotype control PE (553930; BD Pharmingen), and rat IgG isotype control FITC (11‐4031‐81; eBioScience) for 1 h at 4°C in the dark room. The cells were analyzed on FACS Calibur (BD Biosciences) and sorted on an Aria cell sorter (BD Biosciences).

For flow cytometry analyses, the cells were incubated with CD31-PE-Cy^TM7 (PECAM-1); CD45-PE-Cy^TM7 (leukocyte common antigen, Ly-5); CD11b-PE-Cy^TM7 (integrin αM chain); CD34-PE-Cy^TM7 (BD Biosciences, San Jose, CA); CD29-PE, Sca1-APC (Miltenyi Biotec, San Diego, CA); and CD105-PE (eBioscience, San Diego, CA) (1:200) to label hematopoietic, endothelial, and mesenchymal lineages. For cell viability, Sytox Blue 1/1000 (Invitrogen, Thermo Fischer Scientific, Waltham, MA) was added. Analyses were performed using BD LSR Fortessa SORP (BD Biosciences, San Jose, CA). For cell sorting, the cells were resuspended in F10 media (Life Technology, Carlsbad, California) before sorting with BD FACS Aria II SORP (BD Biosciences, San Jose, CA).