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Human peripheral blood mononuclear cells (PBMCs) were incubated with or without (left images) LPS for six hours. After two hours, brefeldin A was added. Cells were harvested, then stained with CD14 antibodies, fixed, permeabilized, and intracellularly stained with Anti-IL-6 antibodies. The FcR Blocking Reagent has been used to avoid Fc receptor–mediated antibody labeling. Cells were analyzed by flow cytometry using the MACSQuant®
Analyzer. Gating was performed on monocytes according to scatter properties of the cells. Autofluorescent cell debris was excluded from the analysis in a fluorescence 4 (FL-4/B3) versus fluorescence 3 (FL-3/B2) dot plot.
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