Cd45ro pe uchl1

FACSAriar (BD Bioscience) was used to perform fluorescent expression analysis. Cells were harvested the following days after transfection and stained with mouse anti-human antibody labeled by fluorescence for 30 m in 4 °C in darks as follows: CD3-PerCP-CY5.5(OKT-3, eBioscience) or CD3-FITC (HIT3a, BD Bioscience), CD4-APC (RPA-T4, BD Bioscience), CD8-PE (HIT8a, BD Bioscience), PD-1-PerCP-CY5.5 (EH12.1, BD Bioscience), CD25-PE (BC96, eBioscience), CD62L-FITC (DREG-56,e Bioscience), CD27-PE (0323, eBioscience), CD28-PE (CD28.2, eBioscience), CD45RO-PE (UCHL1, BD Bioscience), CD69-PE (FN50, BD Biolegend), HLA-DR-PE (G46-6, BD Bioscience).

We performed flow cytometry analysis using the following antibodies: CD3-Percp-Cy5.5 (HIT3a, BD Bioscience), CD8-APC (RPA-T8, BD Bioscience), CD27-PE (M-T271, BD Bioscience), CD28-APC (CD28.2, BD Bioscience), CD45RO-PE (UCHL1, BD Bioscience), CD62L-FITC (DRGE-56, BD Bioscience), αvβ3-FITC (LM6090, EMD Millipore,), αvβ5-FITC (P1F6, EMD Millipore), and NRP-1-PE (AD5-17F6, Miltenyi Biotec GmbH). All samples tested were suspended in FACS buffer and stained with indicated antibodies for 30 min in 4 °C in darks, and then, washed twice, and resuspended in FACS buffer before analysis. For IFN-γ detection, BD™ Cytometric Bead Array (CBA) Human IFN-γ kit (BD Bioscience, USA) was used. Samples were analyzed with BD Accuri C6 (BD Bioscience).