For analysis of surface markers, cells were stained in PBS containing 2% (wt/vol) BSA, with anti-CD4-PB (RM4-5) (Biolegend, 100531, 1:200), anti-CD8α-BV786 (53-6.7) (BD Horizon, 563332, 1:200), anti-TCRβ-APC Cy7 (H57-597) (BD Pharmingen, 560656, 1:200), anti-CD45.2-BV605 (104) (Biolegend, 109841, 1:200), anti-PD-1-APC (RMP1-30) (eBiosciences, 17-9981-82, 1:200), anti-PD-L1-BV711 (10F.9G2) (Biolegend, 124319, 1:100) and anti-CTLA-4-PE (UC10-4B9) (eBiosciences, 12-1522-82, 1:200) on ice for 30 min. Intracellular anti-Foxp3-AF532 (FJK-16 s) (eBiosciences, 58-5773-82, 1:100), anti-IL-2-PerCp Cy5.5 (PC61.5) (eBiosciences, 45-0251-82, 1:100), anti-Ki-67-PE-Cy7 (SolA15) (eBiosciences, 25-5698-82, 1:300), anti-IFN-γ-FITC (XMG1.2) (eBioscience, 11-7311-82, 1:200) and anti-CTLA-4-PE (UC10-4B9) (eBiosciences, 12-1522-82, 1:200), anti-TNF-α-BV650 (MP6-XT22) (Biolegend, 506333, 1:100) were analyzed by flow cytometry according to the manufacturer's instructions. Flow cytometry data were acquired on LSRII or BD FACS Canton II (BD Biosciences) and analyzed using Flowjo software (Tree Star).
Shi L.Z., Fu T., Guan B., Chen J., Blando J.M., Allison J.P., Xiong L., Subudhi S.K., Gao J, & Sharma P. (2016). Interdependent IL-7 and IFN-γ signalling in T-cell controls tumour eradication by combined α-CTLA-4+α-PD-1 therapy. Nature Communications, 7, 12335.
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