Cd19 bv650 hib19

Single cell suspension was processed and stained according to standard protocols. In brief, cell number was adjusted to a concentration of 1–5 × 10⁶ cells/mL in ice-cold FACS Buffer (PBS, 5% FBS, 5 mM EDTA). Then, cells were stained with labeled antibodies for 20 min at 4 °C, washed with FACS buffer and measured. The following antibodies were used: fluorochrome-conjugated monoclonal antibodies against human CD3-APC Cy7 (BioLegend, San Diego, CA, USA), CD5-BV421 (UCHT2, BioLegend, San Diego, CA, USA), CD7-PE (BD Pharmingen, Franklin lakes, NJ, USA), CD19-BV650 (HIB19, BioLegend, San Diego, CA, USA), CD56-APC (BD Pharmingen, Franklin lakes, NJ, USA), CD107a (Invitrogen, Waltham, MA, USA). For intracellular staining, cells were fixed and permeabilized with protein transport inhibitor Golgi Plug (BD, Franklin lakes, NJ, USA) and Monensin (eBioscience, San Diego, CA, USA). Cells were analyzed on a LSR Fortesa (BD Biosciences) using FACSDiva software v.9.0 (Franklin lakes, NJ, USA) Data were analyzed using FlowJo software v.10.7.1 (Tree Star, Ashland, OR, USA).