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Mouse spleen cells were incubated with or without KLH for 6 hours. After 2 hours, brefeldin A was added. Cells were harvested, fixed, permeabilized, and intracellularly stained with Anti-IFN-γ antibodies and CD154 antibodies. Cells were analyzed by flow cytometry. Gating was performed according to CD4 expression and side scatter properties of the cells.
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