Ter119 ly 76

For assessment of bone marrow and spleen progenitor cells by flow cytometry, single-cell suspensions of freshly prepared bone marrow and spleen were incubated with the following anti-mouse monoclonal antibodies: Ter119 (Ly-76, BD Biosciences), CD71 (C2, BD Biosciences), CD34 (RAM34, BD Biosciences), CD135 (A2F10, eBioscience), CD127 (A7R34, eBioscience), c-Kit (ACK2, eBioscience), SCA-1 (LY-6A/E, BD Biosciences), eFluor520 (eBioscience), and 7-AAD (eBioscience). Lineage (lin) staining included anti-mouse Ter119 (BD Biosciences), GR-1 (RB6-8C5, BD Biosciences), B220 (RA3-6B2, BD Biosciences), CD4 (RM4-5, BD Biosciences), CD8 (53–6.7, BD Biosciences), and CD11b (M1/70, BD Biosciences) antibodies [22 (link)–24 (link)]. Multicolor data acquisition for bone marrow progenitor subsets was performed on a FACSCanto II flow cytometer, and data were analyzed with FlowJo 7.6.1.

Flow cytometry was performed on a BD LSRI or LSRII (BD Biosciences, San Jose, CA), and data were analyzed using FlowJo software (TreeStar, Ashland, OR). Antibodies include CD45.2, CD45.1, Ly-6G (Gr-1), CD11b (Mac-1), CD45R/B220, CD4 (L3T4), CD8 (Ly2), and Ter119/Ly76, Sca1 (Ly-6A/E), c-Kit (CD117), CD34, CD16/32, IL7Ra (BD Bioscience).
BD Aria was used for cell sorting. BM cells were lineage depleted using a lineage depletion kit (Miltenyi Biotec, Auburn, CA), and labeled with phycoerythrin (PE)-conjugated lineage antibodies, fluorescein isothiocyanate (FITC)-conjugated Sca-1 and allophycocyanin (APC)-conjugated c-Kit, LSK cells were sorted.