Figure 2.

Gating strategy. Cells were gated side scatter area vs forward scatter area then Zombie Aqua dye (which penetrates non-viable cells but not viable cells; Biolegend, San Diego, USA) was used to gate on live cells only. After that, cells were gated on side scatter height vs side scatter area to exclude duplicate cells. Cells were then gated on CD3 marker and then on CD4 and CD8 markers. Lastly, cells were gated based on marker of interest (CD69, FasR, CD137 and CD107a) as shown on the following figures. The arrows indicate stepwise progression through each of the gating steps.