FIGURE 1.

Basic gating strategy for flow cytometry experiments. A side-scatter (SSC) vs. forward scatter (FSC) plot with all the events recorded. Cells were selected “All” after size-based exclusion of cellular debris (A). Forward scatter- height vs. area plot allowed for selection of “singlets,” avoiding cells sticking to each other as doublets (B). “Viable/live” cells (zombie aqua negative) were then positively gated from the singlets (C). From this live cell pool, CD45⁺ “immune” cells were gated (D). From the immune cells, CD3⁺ “T cells” were selected. From this CD3+ cells, CD4/CD8 T cells and their subpopulations were further gated (E). Then from the CD3^– cells, B220 expressing cells were gated as B cells. B220⁺CD25⁺ cells were further gated from B cells. CD3^–B220^– cells were further gated into NK cells or myeloid cells and their subtypes (F).