Intracellular ATP delivery to in vitro expanded mouse CD27− γδ T cells

Intracellular ATP supports the function of γδT17 cells in mice. Here, we present a protocol for intracellular ATP delivery to in vitro expanded mouse CD27 γδ T cells. We describe steps for pre-coating well plates, preparing lymphocytes, culturing CD27 γδ T cells, and ATP delivery. We then detail functional evaluation of γδ T cells by flow cytometry. Appropriate concentrations of control and ATP vesicles are detailed for intracellular ATP delivery, which can also be applied to other immune cells. For complete details on the use and execution of this protocol, please refer to Wang et al. (2023)..