2.5. Flow Cytometry
Cells were stimulated with Phorbol 12-Myristate 13-Acetate (PMA),
Ionomycin, Golgi Stop (50 ng / ml, 1 µg / ml, 1 µl / ml, respectively)
for 4 hours in a 37ᵒC incubator. Cells were fixed for 15 minutes and
permeabilized with the BD Cytofix / Cytoperm ™ Plus kit. The single cell
suspension was stained with antibodies 30 min on ice at dark after 5
minutes after blocking with mouse TruStainFcX (BioLegend, San Diego, CA,
USA) in staining buffer (2% FBS in PBS) with appropriate dilution. Data
collection was performed on FacsAriaIII. FlowJo or Diva software was
used to analyze flow cytometry data.