“…Cells were then washed twice in 10 ml of HBSS + 10% FBS and centrifuged at 600 rcf for 3 minutes and resuspended in complete RPMI. For sorting, cells were washed in PBS twice and stained with the following antibodies: KLRG1 FITC (2F1; eBioscience 11-5893-82), CD62L Percp-cy5.5 (MEL-14; eBioscience 45-0621-82), CD8a PE-cy7 (53–6.7; BioLegend 100722), CD127 BV421 (A7R34; BioLegend 135024),Viability-Zombie Aqua (BioLegend 423101), CD44 APC (IM7; eBioscience 17-0441-82), CD4 BV711 (RM4-5; BioLegend 100550), PD-1 APC-750/Fire (29F.1A12; BioLegend 135240), and mouse NRP-V7 mimotope tetramer PE (KYNKANVFL/H-2K d ; NIH tetramer facility) 30 . Cells were sorted on viability dye-negative, CD4 − CD8 + , and tetramer + CD44 + on a Sony ICyt Synergy (Sony Biotechnology, Inc.).…”