Pe cyanine7 anti human cd8a

Cells were collected and washed 2 to 3 times. The cell surface mixed antibody was divided equally into each sample staining with the Zombie aqua fixable viability kit (423101, BioLegend), and continued simultaneously for 30 min at room temperature. After the cell surface staining was finished, the intracellular maker staining was performed. Cell fixation (420801, BioLegend) and staining permeabilization (421002, BioLegend) were required prior to intracellular staining. Flow cytometry determined the expression of distinct surface and intracellular molecules. The following antibodies were used: APC/Cyanine7 anti-human CD3 (300317, BioLegend), PerCP/Cyanine 5.5 anti-human CD4 (317427, BioLegend), PE/cyanine7 anti-human CD8a (300913, BioLegend), Brilliant Violet 421 anti-human IFN-γ (502531, BioLegend), and PE/DazzleTM594 anti-human CD107a (LAMP-1) (328645, BioLegend) and APC anti-human TNF-α (502913, BioLegend), APC anti-human CD19 (302211, BioLegend). The apoptosis of BJAB cells was assayed using the FITC Annexin V Apoptosis Detection Kit with PI (640914, BioLegend). The data were analyzedusing FlowJo 10.8 software (https://www.flowjo.com/).