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T cells were enriched with the CD4+
T Cell Isolation Kit II, mouse from splenocytes of BALB/c mice. Cells were stimulated with plate-bound CD3 and soluble CD28 antibodies in the presence recombinant IL-4, IL-2, TGFß-1 and neutralizing Anti-IL-6 antibodies, Anti- IFN-γ antibodies and Anti-IL-12 antibodies for 5 days. Cells were harvested, fixed, permeabilized, and intracellularly stained with Anti-IL-9 antibodies or with the REA Control antibodies (left image), as well as with CD4 antibodies. Cells were analyzed by flow cytometry using the MACSQuant®
Analyzer. Cell debris were excluded from the analysis based on scatter signals.
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