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A mixture of lymphnodes and splenocytes from C57BL/6 mice was isolated with the Naive CD4+
T Cell Isolation Kit. The separated cells were then Tʜ17 polarized for 3 days at 37 °C. After that cells were restimulated with 500 ng/mL PdBu and ionomycin and 1 µg/mL brefeldin A. Cells were fixed, permeabilized, and stained with Anti-IL-17A antibodies as well as with CD4 antibodies. Cells without restimulation are shown in left image. Flow cytometry was performed using the MACSQuant®
Analyzer. Cell debris were excluded from the analysis based on scatter signals.
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