Cd161 apc

PBMC were stained with the following antibodies: TCRC Pacific Blue, CD4 V500, V follow (Biolegend, UK), MR1 APC (26.5) (Biolegend, UK), CD3 PerCP, CD161 APC, CD8 APC-Cy7, CD19 APC-Cy7 and CD20 Pacific Blue. All antibodies were from BD biosciences unless specified. The samples were stained for 15 minutes at room temperature, processed and then analyzed on a FACS Canto II machine with FACS Diva software evaluation.

All flow cytometry and analysis was done on an Aria FACS Diva flow cytometer equipped with 3 lasers, and all analysis was done with the FACS Diva software. The following antibodies were used in specific phenotypic panels: T-Cell Panel: CD4-APC-Cy7 (Biolegend, San Diego, CA, USA), CD8a-V450 (BD Horizon San Jose, CA, USA), CD44-FITC (ThermoFisher Scientific, Waltham, MA, USA), CD62L-eFluor660 (eBiosciences, San Diego, CA, USA), KLRG1-Biotin (BD Pharmingen, San Jose, CA, USA) with Streptavadin-Alexa Fluor 430 (ThermoFisher Scientific) as the secondary, and CD127-PE (R&D Systems, Minneapolis, MN, USA). NK Cell Panel: CD56-Alexa Fluor 700 (Novus Biologicals, Littleton, CO, USA), CD314-PE (ThermoFisher Scientific), CD3-FITC, CD161-APC, CD8a-PerCP all from Biolegend. DC Panel: CD11c-FITC (Bio-Rad, Hercules, CA, USA), CD86-PE (Biolegend), and MHC Class II-APC (Miltenyi Biotec, Teterow GER). Treg Panel: FoxP3-PE, CD4-APC and CD25-FITC all from Biolegend. All antibodies were used at the company’s suggested concentration or titrated for optimal use.