Analysis of Tim-3 interactome in primary mouse CD4+ T cells

We analyzed the interactome of the Tim-3 (HAVCR2) protein, using an affinity purification method based on the the endogenous expression of a One-Strep-tagged (OST) version of Tim-3 in an engineered mouse model, and affinity purification of the protein from expanded primary CD4+ T cells. Affinity purification of the OST tagged protein was performed using Streptactin beads, from T cells either non stimulated, or stimulated 30s, 120s, 300s or 900s with pervanadate. Each AP-MS purification is associated with a corresponding control (purification from CD4+ T cells isolated from WT mice, cultured and stimulated in the same conditions). The number of replicate biological experiments was n=3 for all conditions (time-points), and each sample was analyzed twice by nanoLC-MS, resulting in 60 raw files.