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).

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.