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Human peripheral blood mononuclear cells (PBMCs) were either left unstimulated (left image) or stimulated with 20 ng/mL phorbol 12-myristate 13-acetate (PMA) and 1 µg/mL ionomycin for 2 hours, followed by an incubation with 1 µg/mL brefeldin A for 4 hours. Cells were fixed, permeabilized, and directly stained with Anti-IL-3 antibodies as well as with CD154 antibodies. Flow cytometry was performed using the MACSQuant®
cells were pregated for the analysis. Cell debris were excluded from the analysis based on scatter signals.