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Splenocytes from C57BL/6 mice were either left unstimulated (left images) or stimulated with plate-bound CD3 antibodies and soluble CD28 antibodies for 24 hours. Cells were then stained with CD153 antibodies as well as with CD4 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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