For the in vitro killing assay 50,000 MC38-GFP cells were stained with CellTrace Violet (Invitrogen, Thermo Fisher Scientific), mixed with 50,000 MC-38-OVA-GFP cells stained with CellTrace Yellow (Invitrogen, Thermo Fisher Scientific), and incubated for 24 hours at 37°C. Cells were then kept alone or cocultured with enriched CD8+ T cells from spleens of either Ptpn2fl/fl Cd4Cre– OT I or Ptpn2fl/fl Cd4Cre+/– OT I mice for 24 hours in a 37°C incubator. CD8+ T cells were enriched using the EasySep CD8+ T Cell Enrichment Kit (STEMCELL Technologies). Apoptosis was then detected using the Annexin V Kit – DY-634 Antibody (Abcam) and the Zombie NIR Fixable Viability Kit (BioLegend).

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