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Mouse spleen cells were either left unstimulated (left peak) or stimulated with plate-bound CD3 and soluble CD28 antibodies for 3 days. After that cells were stained with CD137 antibodies and analyzed by flow cytometry using the MACSQuant®
Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.
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