Ly6g 6c

Bone marrow cells, thymocytes, splenocytes, or peripheral blood cells were isolated, and RBC lysis buffer was added to remove the RBCs. Cells were stained at a 1:200 dilution with 15 mouse fluorochrome-conjugated monoclonal antibodies specific for the following murine cell surface markers encompassing the major immune lineages: B220, CD19, IgM, IgD, CD3ε, CD4, CD5, CD11c, CD44, CD43, CD25, CD21, CD23, BP-1 (BD PharMingen), CD8α, CD11b, NK1.1 (Biolegend), F4/80, and CD62L (Tonbo Biosciences), and in the presence of anti-mouse CD16/32 antibody (Tonbo Biosciences) for 1 h at 4°C. After staining, cells were washed twice in PBS and analyzed by flow cytometry. To stain the hematopoietic progenitor compartment, bone marrow was isolated and stained with Alexa Fluor 700–conjugated lineage markers (CD3, Ly-6G/6C, CD11b, B220, and Ter-119 at a 1:50 dilution; Biolegend), c-Kit, Sca-1, CD16/32, CD34, IL-7Rα (BD PharMingen), and CD135 (Biolegend) at a 1:100 dilution for 1 h at 4°C. After staining, cells were washed twice in PBS and analyzed by flow cytometry. Data were acquired on an LSRFortessa cell analyzer (BD Bioscience) and analyzed with FlowJo software (Treestar).