Gating strategy for cultured cells. A, Representative FlowJo cytometry plots show that forward scatter (FSC) versus side scatter (SSC) was used to draw a gate around the cells. B, Single cells were gated by plotting FSC height against FSC area to exclude doublets. C, Viable cells were gated by excluding dead cells stained with Ef780 dye. D, Single viable cells were then analyzed for individual markers, including HLA-DR (lighter line), overlaid with their isotype controls. See Table E1 for full phenotypic details.