Efluor 506 ebioscience

Samples were stained with fixable viability dye eFluor 506 eBioscience (Thermo Fisher Scientific, Waltham, MA) and fluorophore-conjugated antibodies against mouse CD16/32, CD45, CD45.1, CD45.2, CD45R/B220, CD3, CD19, NK1.1, Ly6G, PDCA-1, MHC-II, CD11b, CD11c, CD103, CD115, CD117, CD127, CD135, Sca-1, Gr-1, and TER-119 (Supplementary Table 1). Unless stated otherwise, all cell populations gated on live single cells and identified as follow: leukocytes: CD45^pos; plasmacytoid dendritic cells (pDCs): CD45^poslineage(CD3,CD19, NK1.1)^negLy6G^negCD11c^posPDCA-1^pos; classical dendritic cells (cDCs): CD45^poslineage(CD3,CD19,NK1.1)^negLy6G^negPDCA-1^negMHC-II^hiCD11c^hi; macrophage/dendritic cells progenitor (MDP) and common dendritic cell progenitor (CDP) were both identified as: CD11b^negTER119^negB220^negCD3^negGr1^negSca1^negCD16/32^posCD135^pos, then MDP identified as: CD115^posCD117^pos and CDP identified as: CD115^posCD117^neg. Common lymphoid progenitor (CLP) was identified as: CD11b^neg− TER119^negB220^negCD3^negGr1^negSca1^posCD117^posCD127^posCD135^pos. Samples were acquired using an LSRFortessa flow cytometer (BD Bioscience, San Jose, CA). FlowJo software version 10.0.8r1 (FlowJo, LLC. Becton, Dickinson and Company, Franklin Lakes, NJ) was used for analysis of the acquired flow cytometry data.