Extended Data Fig. 1: Gating Strategy.
From: β-Glucan reprograms neutrophils to promote disease tolerance against influenza A virus
(a) Cells were gated for FSC-A against SSC-A. Doublets were excluded using FSC-H against FSC-A. Viable CD45+ cells were gated, and within the viable CD45+ cells, cells were gated as CD11b, excluding SiglecF+ cells. CD11b and Ly6G double-positive cells were gated as neutrophils. (b) Cells were gated for FSC-A against SSC-A and doublets were excluded using FSCH against FSC-A as shown in (a). Viable cells were gated, and lineage-committed cells were excluded. Within the lineage-negative population, cells were gated as CD127+ and CD127-. Lin-CD127- population was further gated as LKS-defined as double positive for cKit and Sca-1, and cKit is gated as Sca-1 negative and cKit positive. Gated on the LKS population, cells were divided into LT-HSC, STHSC and MPP based on CD150 and CD48 expression. C-Kit+ Sca-1- cells were further gated based on CD34 and CD16/32 to define CMP, GMP and MEP. (c) Finally, in another set of experiments, Lineage+ cells and then Sca-1+ cells were excluded. The remaining cells were subdivided into cKit+ CD16/32+ (II) and cKit+ CD16/32- groups. In the cKit+CD16/32 + , cells were further gated on CD34+ Flt3- cells. Within this fraction, Ly6C + CD115- cells were the GP, and Ly6C + CD115+ were cMoP.
