Flow cytometric analysis was performed on an LSR Fortessa (BD Biosciences) and data analyzed using FlowJo Version 10.7.0 (Treestar). The following anti-human antibodies were used: anti-CD3-PE-Cy7 (BD Biosciences, clone HIT3a), anti-CD4-V500 (BD Biosciences, clone RPA-T4), anti-CD25-APC (Biolegend, clone BC96), anti-CD152-PE (CTLA-4) (BD Biosciences, clone BNI3), LIVE/DEAD TM Fixable Near-IR dead cell stain kit (Thermo Fisher Scientific, L10119). Intracellular staining was performed using the eBioscience TM Foxp3/Transcription factor staining buffer set (Thermo Fisher Scientific, 00-5523-00) and anti-FOXP3-APC (Thermo Fisher Scientific, clone 236A/E7) and anti-GFP-FITC (Rockland, 600-402-215).
The following anti-mouse antibodies were used: anti-CD3-BV421 (BD Biosciences, clone 17A2), anti-CD4-BUV737 (BD Biosciences, clone GK1.5), anti-CD4-PerCP-Cy5.5 (BD Biosciences, clone RM4-5), anti-CD152-PE (CTLA-4) (BD Biosciences, clone UC10-4F10-11), anti-FOXP3-APC (Thermo Fisher Scientific, FJK-16s), Fixable Viability Dye eFluorâ„¢ 780 (Thermo Fisher Scientific).
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