Fitc conjugated anti mouse foxp3

For cell surface and intracellular marker analysis, 10⁶ cells were incubated with fluorescent-conjugated antibodies in labeling solution (eBioscience, USA). The fluorescent-conjugated antibodies used in this study included PE-conjugated anti-mouse CD11c, FITC-conjugated anti-mouse CD11c, FITC-conjugated anti-mouse CD86, FITC-conjugated anti-mouse MHC-II, APC-conjugated anti-mouse CD80, PE-CY5-conjugated anti-mouse CD40, APC-conjugated anti-mouse CD4, FITC-conjugated anti-mouse IL-17A, FITC-conjugated anti-mouse Foxp3, PE-conjugated anti-mouse GATA-3, PE-conjugated anti-mouse T-bet antibodies, and FITC-conjugated anti-mouse CD103 (integrin alpha E) (all purchased from eBioscience, USA). Fluorescent-conjugated, isotype-matched, irrelevant antibodies were used to establish background fluorescein levels. Flow cytometry analysis was conducted on a FACSCalibur (BD Biosciences, USA), and FACSCalibur software (BD Biosciences) was used to analyze the flow data. DCs were gated for PE-CD11c, and then FITC-MHC-II expression and endocytic FITC-OVA levels were analyzed. MLR-lymphocytes were gated for APC-CD4, and then FITC-Foxp3 was analyzed.