CHIP-seq of healthy donor-derived CD8+ T cells in flask and 3D conditions

Cells were then collected by incubating flask and 3D fibrin gel for 3 days. ChIP was performed by using an iDeal ChIP-seq kit for Histones (Diagenode) according to the manufacturer’s protocol.

In brief, 1×107cells were crosslined with 1% formaldehyde for 8 min at room temperature. After stopping the fixation with 0.125M glycine for 5 min at room temperature, cells were washed, lysed and sheared by sonication using Bioruptor (Diagenode) for 20-30 cycles (30s “ON”, 30s“OFF”) at high power setting. The sheared chromatin fragments were immunoprecipitated with protein A-coated magnetic beads and anti-P73; anti-RUNX2; anti-TEAD or anti-IgG (Cell Signaling, Cat.: 3900S; 1:50) antibody.