Whilst FACS cannot provide important information about tissue location of individual immune subsets in relation to the tumor, FACS did provide additional information indicating that TILs comprised largely effector memory T cells, including activated CD8+ and CD4+ T cells which expressed PD-1 immune checkpoint (and not TIM-3), PD-1+ and IL-7Rα+ CD8+ T cells were mutually exclusive, and OX-40 was only present on CD4+ T cells. Here, TNFRSF4 is linked to neoplasm.