Ly6g bv605

One million live cells were counted using trypan blue and first stained with a fixable viability dye (eBiosciences 65-0866-14, Thermo Fisher Scientific; Waltham, MA, USA) for 30 min at 4 °C. Cells were washed in flow wash buffer (PBS with 0.05% sodium azide and 3% FBS) and stained with surface antibodies for 40 min at 4 °C. Cells were washed in a flow buffer and fixed in a flow buffer plus 4% PFA before filtering through 40-μm mesh strainer tubes (BD Biosciences; San Jose, CA, USA). Flow cytometry was performed on the BD Fortessa X20 cytometer, and the data were analyzed using FlowJo Version 10 (Becton, Dickinson & Co.; Franklin Lakes, NJ, USA). Flow cytometry antibodies used from BD Biosciences (San Jose, CA, USA) include: CD45-APC-R700 (565478), CD8-PE (553033), CD4-APC-H7 (5560181), Ly6G-BV605 (563005), Ly6C-FITC (561085), CD11b-APC (561690), PD-1-BV421 (562584), NK1.1-BV650 (564143), and MHCII-PerCPCy5.5 (562363). Flow cytometry antibodies used from eBiosciences (Thermo Fisher Scientific; Waltham, MA, USA) include: F4/80-Super Bright 780 (78-4801-82).