Cd34 percp eflour710

FLSs were blocked with 0.1 mg/mL mouse IgG (Jackson ImmunoReseach, West Grove, PA, USA) and 0.1 mg/mL rat IgG (Jackson ImmunoReseach) for 15 min and then surface stained with CD34-PerCP-eFlour710 (Clone: 4H11, Thermo Fisher Scientific, Waltham, MA, USA), CD45-APC-Cy7 (Clone: HI3, Biolegend), THY-1-PE-Cy7 (Clone: 5E10, Biolegend, San Diego, CA, USA), PDPN-PE (Clone: NZ-1.3, eBioscience, Thermo Fisher), and fixable LIVE/DEAD nIR Dead Cell Stain Kit (Life Technologies, Thermo Fisher Scientific). Data were acquired by NovoCyte Quanteon^® (ACEA Bioscience Inc., San Diego, CA, USA) within 24 h of surface stain and processed in FlowJo (FlowJo software version 10.5.3). The spectral overlap was compensated using antibody-coated beads (eBioscience, San Diego, CA, USA). Gating was performed on live cells using fluorescence minus one (FMO).

Fibroblasts were blocked with 0.1 mg/mL mouse IgG (Jackson ImmunoReseach) and 0.1 mg/mL rat IgG (Jackson ImmunoReseach) for 15 min and stained with CD34-PerCP-eFlour710 (Clone: 4H11, Thermo Fisher Scientific), CD45-APC-Cy7 (Clone: HI3, Biolegend, San Diego, CA, USA), THY-1-PE-Cy7 (Clone: 5E10, Biolegend), Podoplanin-PE (Clone: NZ-1.3, eBioscience, Thermo Fisher Scientific), ICAM-1-BV421 (Clone: HA58, BD Bioscience, Franklin Lakes, NJ, USA), HLA-DR-BV650 (Clone: L243, Biolegend) and fixable LIVE/DEAD nIR Dead Cell Stain Kit (Life Technologies, Thermo Fisher Scientific). Data were acquired on NovoCyte Quanteon (ACEA Bioscience inc., San Diego, CA, USA) and processed in FlowJo (FlowJo software version 10.5.3).