Figure 3.

Sorting strategy for Nanostring arrays. Lymphocytes were identified by forward -scatter area (FSC-A) and side scatter area (SSC-A) gates, followed by two singlets gates (FSC-A vs. FSC-W and SSC-A vs. SSC-W) followed by live/dead identification using the infra-red fluorescent viability dye. Alive CD45⁺ cells were plotted for CD11b⁺ vs dump channel (anti-NK1.1/Ly6G/CD3e/CD45R/B220) to remove NK cell, granulocyte, T cell and B cells lineages, respectively. The CD11b⁺ cells were further gated on MHCII. The MHCII^- population was sub-gated on Ly6C as MHCII^-CD64⁺Ly6c^high (monocytes), MHCII^-CD64⁺Ly6c^inter and MHCII^-CD64⁺Ly6c^low (macrophages). Cells were sorted from twenty C57BL/6 mice (n = 20) to obtain sufficient RNA to perform the NanoString expression profiles.