Fig. 6

Gating strategies for the surface marker analysis of human peripheral blood mononuclear cells (PBMCs) (a) and macrophages (b). a PBMCs were first gated (black lined gate) on via sideward scatter area (SSC-A) and forward scatter area (FSC-A) on lymphocytes. Doublets were excluded by gating on forward scatter height (FSC-H) vs. FSC-A and then viable cells were gated by FSC-A vs. Live/Dead Marker. Thereafter, CD3 + CD45 + T cells were identified based on gating by CD3 vs. CD45, which were then separated into CD4 + and CD8 + T cells. Finally, the mean fluorescence intensity (MFI) of the marker of interest (filled grey histogram) was determined in comparison to the negative control staining (black lined histogram). b Macrophages were first gated via SSC-A vs. FSC-A to exclude cell debris from the analysis. Doublets were excluded by gating on FSC-H vs. FSC-A. Then, viable cells were identified (FSC-A vs. Live/Dead) and the MFI of the marker of interest (filled grey histogram) was determined in comparison to the negative control staining (black lined histogram)