CD107a Degranulation Assay

T cells (3 × 10⁵/well) on days 13–14 after transduction were incubated with an equal number of target cells treated with the EZH2 inhibitor for 14 days in a total volume of 200 μL, in Eppendorf tubes in the presence of Monensin (eBioscience, Germany) (1 μL/mL) and CD107a-PE antibody (BioLegend, Germany) (100 ng/mL) for 3 h at 37°C and 5% CO₂. The cells were washed, then incubated with CF488-conjugated G_D2 anti-idiotype antibody ganglidiomab and a CD3-specific antibody for 15 min, fixed in 1% PFA, and analyzed by flow cytometry. T cells incubated with medium alone or with DMSO-treated EwS cells served as controls.