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Human peripheral blood mononuclear cells (PBMCs) were either left unstimulated (left peak) or stimulated with 760 nM Phorbol 12-myristate 13-acetate (PMA) at 37 °C for 15 minutes. Cells were then fixed and permeabilized using the Cell Signaling Buffer Set A followed by intracellular staining with Anti-S6 pS235/pS236 antibodies. Flow cytometry was performed with the MACSQuant®
Analyzer. Cell debris were excluded from the analysis based on scatter signals.
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