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C57BL/6 splenocytes were stained with CD196 (CCR6) antibodies as well as with CD45R (B220) antibodies and analyzed by flow cytometry using the MACSQuant®
cells were pregated for the analysis. 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.