Anti mouse cd19 apc

Isolated immune cells were analyzed by staining fluorescence conjugated antibodies against cell surface markers and intracellular proteins. Anti-mouse CD4-APC, anti-mouse CD19-APC, anti-mouse H-2Dd-BV421, anti-mouse H-2Db-FITC, anti-mouse CD8a-PE, anti-mouse NK-1.1-PE-Cy7, and anti-mouse IFN-γ-PerCP-Cy5.5 antibodies were purchased from BD Biosciences (San Jose, CA). Anti-mouse CD11c-APC, anti-mouse FoxP3-PerCP, and anti-mouse CD25-PE antibodies were purchased from eBioscience (San Diego, CA). Anti-mouse CD11b-PE antibody was purchased from Miltenyi Biotec (Bergisch Gladbach, Germany). For Treg cell staining, immune cells were first stained with cell surface markers CD4 and CD25 and fixed overnight at 4°C, then permeabilized for 30 min at room temperature, and finally stained with intracellular FoxP3 for 30 min. For T helper cell intracellular cytokine staining (Th1, Th17, and Th2 cells), immune cells were first activated in the presence of phorbol 12-myristate 13-acetate (20 ng/mL), ionomycin (1 μg/mL), and monesine (4 μM) for 4 h, and then staining of cell surface and intracellular cytokines was carried out.