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Human peripheral blood mononuclear cells (PBMCs) were either left unstimulated (left images) or stimulated with 50 ng/mL PMA and 1 µg/mL ionomycin for 6 hours. After 2 hours, 1 µg/mL brefeldin A was added. Cells were then fixed, permeabilized, and intracellularly stained with CD152 antibodies as well as with CD4 antibodies. Flow cytometry was performed using the MACSQuant®
Analyzer. Cell debris were excluded from the analysis based on scatter signals.