Cd45.2 bv421

The following antibodies were used: mouse lineage cocktail-PE (BioLegend, cat# 78035), mouse lineage cocktail-APC (R&D Systems, cat# FLC001A), c-Kit-FITC (BioLegend, cat# 161603), c-Kit-APC (BioLegend, cat# 135108), c-Kit-PE/Cy7 (BioLegend, cat# 105814), c-Kit-BV421 (BioLegend, cat# 135124), Sca-1-BV605 (BioLegend, cat# 108133), Sca-1-APC (eBioscience, cat# 17-5981-82), Sca-1-BV421 (BioLegend, cat# 108127), Sca-1-PerCP-Cy5.5 (eBioscience, cat# 45-5981-82), CD150-BV421 (BioLegend, cat# 115925), CD150-PE (eBioscience, cat# 12-1501-82), CD150-PE-Cy7 (BioLegend, cat# 115913), CD48-PE/Cy7 (eBioscience, cat# 25-0481-80), CD48-BV711 (BioLegend, cat# 103439), CD48-APC-Cy7 (BioLegend, cat# 103432),CD34-FITC (eBioscience, cat# 11-0341-82), CD135-PE-Cy5 (BioLegend, cat# 135311), CD135-APC (BioLegend, cat# 135310), CD45.1-PerCP-Cy5.5 (BioLegend, cat# 110727), CD45.2-BV421 (BioLegend, cat# 109831), CD45.2-FITC(eBioscience, cat# 11-0454-82), Gr1-PE (BioLegend, cat# 108407), CD11b-APC (BioLegend cat# 101211), CD4-PE (BioLegend,cat# 116005), CD8a-PE (BioLegend, cat# 100707), B220-APC (BioLegend, cat# 103211).

Primary KCs and ImKCs were immunostained for surface markers. Fixable Viability Dye eFluor 780 (1:1000, #65-0865), CD11b BV421 (1:75, clone M1/70, #404-0112-80) and TIM-4 PerCP eFlour 710 (1:75, clone RMT4-54, #12-5866-82) were purchased from Thermo Fisher (Thermo Fisher Scientific, Waltham, MA, USA). CD45.2 PE (1:5, clone 104, #109808), CD45.2 BV421 (1:75, clone 104, #109831), F4/80 APC (1:75, clone BM8, #123116), CLEC4F AF647 (1:75, clone 3E3F9, #156803), CLEC2 PE (1:75, clone 17D9/CLEC-2, #146103), CD14 PE (1:75, clone Sa-14-2, #123309), TLR4 APC (1:75, clone SA15-21, #145405) were purchased from BioLegend (San Diego, CA, USA). Unstained cells as well as fluorescent minus one (FMO) control samples were used as controls on every staining. All stainings were performed in the presence of the Fc receptor’s blocker monoclonal antibody (Bio X Cell, Lebanon, NH, USA, 20 μg/mL, clone 2.4G2, #BE0307). Samples were measured on a CytoFLEX cytometer (Beckman Coulter, Brea, CA, USA). Data were analyzed using the FlowJo software v10.8.2 (BD Biosciences, Franklin Lakes, NJ, USA).