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Human peripheral blood mononuclear cells (PBMCs) were either left unstimulated (left images) or were stimulated with CytoStim™ for six hours. After two hours, brefeldin A was added to stimulated and unstimulated cells. The cells were fixed, permeabilized, and intracellularly stained with CD154 antibodies as well as with CD4 antibodies and analyzed by flow cytometry using the MACSQuant®
Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye–conjugated antibodies. CD4+
lymphocytes were pre-gated for the analysis. Cell debris were excluded from the analysis based on scatter signals.
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