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Jurkat human PD1-CD3ζ NFAT-GFP reporter. (a) Kozak-huPD1-CD3zeta sequence. The human PD1 extracellular and transmembrane domain were coupled to the human CD3ζ signaling domain (everything after the transmembrane domain). (b) Jurkat PD1-CD3ζ NFAT-GFP reporter and NFAT-GFP reporter control. PMA/ionomycin stimulation results in a maximum GFP signal of 20–30%, while PD-L1–Fc only achieves 5% GFP-positive cells. The system is sub-optimal but can detect PD1–PD-L1 interaction.
