Cd56 pc5

For flow cytometry, cells were washed with PBA [PBS supplemented with 0.5% bovine serum albumin (BSA), 1% FBS and 0.1% sodium azide] and incubated with antibodies against surface markers [CD3-CF (Immunostep); CD3-FITC, CD56-PC5, CD16-PE-Cy7, CD4-PC5.5, CD8-PC7 and LAMP1-APC (Biolegend)] at 4°C for 30 min in the dark. For intracellular staining, after surface labelling, cells were fixed with 1% p-formaldehyde for 10 min at RT, permeabilized with 0.2% saponin for 10 min at RT and stained with Perforin-PE, Granzyme B-PE (Biolegend) for 30 min. Cells were washed in PBA and analyzed using a Gallios Flow Cytometer (Beckman Coulter). Analysis of the experiments was performed using Kaluza software.

For the flow cytometry, the cells were washed with PBA (PBS supplemented with 0.5% bovine serum albumin (BSA), 1% FBS, and 0.1% sodium azide), and incubated with conjugated antibodies against surface markers (CD3-CF (Immunostep, Salamanca, Spain); CD56-PC5 (Phycoerythrincyanin 5), CD16-PE-Cy7 (phycoerythrin-Cy7), CD14-APC (Allophycocyanin), CD15-PB (Pacific Blue), and CXCR3-APC (BioLegend, San Diego, California, USA) at 4 °C for 30 minutes in the dark. For intracellular staining, after surface labelling, the cells were fixed with 1% p-formaldehyde for 10 min at room temperature (RT), permeabilized with 0.2% saponin for 10 min at RT, and stained with anti-CXCL10-PE (BD, San Jose, California, USA) or mouse IgG2a-PE (Beckman Coulter, Brea, California, USA) for 30 min. The cells were washed in PBA and analyzed using a Gallios Flow Cytometer (Beckman Coulter, Brea, California, USA). The analysis of the experiments was performed using either Kaluza (Beckman Coulter, Brea, California, USA) or FlowJo software (Ashland, Oregon, USA, https://www.flowjo.com).