Cd183

T cells from in vitro primary murine T cell culture, fresh peripheral blood, or whole splenocytes were analyzed according to cell surface and intracellular markers. Surface markers were directly stained with flow antibodies for 20 min at 4 °C. For intracellular staining (FOXP3 and T-BET), cells were fixed and permeabilized after surface staining using appropriate buffers (One-step intracellular nuclear protein staining protocol, eBioscience, San Diego, CA, USA) and stained with flow antibodies. The antibodies used for flow cytometric analyses for mouse cells were as follows: FOXP3, T-BET, and CD183 (eBioscience, San Diego, CA, USA); CD3, H2-Kb, H2-Kd, CD4, B220, CD11b, CD11c, Ly6C, and DEC205 (BioLegend, San Diego, CA); and CD8, CD45.2, CD80, and Ly6G (BD Pharmingen, San Jose, CA, USA). All cells were run on a ZE5 Cell Analyzer (BD Biosciences, Mountain View, CA, USA) or Attune^® NxT (Thermofisher Scientific, Waltham, MA, USA) and analyzed with the FlowJo software, version 10.9.0 (FlowJo LLC., Ashland, OR, USA).