GSM6751674
The cells were then crosslinked with fresh 1% formaldehyde (Thermo Fisher Scientific) for 10 min at room temperature (RT) and quenched with 125 mM glycine for 5 min at RT. Cells were washed three times in cold PBS containing protease inhibitors. Nuclei were isolated using the SimpleChIP Enzymatic Chromatin IP Kit (#91820, CST), and chromatin was sheared on ice using a Fisher sonic dismembrator with the setting of 10 cycles of 4 sec “on” and 8 sec “off”. 2% input fraction was saved, and the remaining sheared chromatin was used for ChIP with a rabbit anti-Xbp1s antibody (Cat No. 40435, CST) or rabbit anti-T-bet antibody (97135, CST) or an IgG isotype control (Millipore) in 1 × ChIP buffer with rotating overnight at 4°C. Protein G magnetic beads were added to each immunoprecipitation and incubated for 2 h at 4°C with rotation. The chromatin bead antibody complexes were then washed three times with low salt wash buffer and one-time with high salt wash buffer. Chromatin was then eluted using 1% SDS in tris-EDTA. Cross-linking was reversed by incubation with 5M NaCl and Proteinase K at 65°C overnight. DNA was purified with ChIP DNA Clean & Concentrator kit (Zymo Research). ChIP-Seq libraries were prepared using kapa Hyper DNA kit (Roche) and sequenced on an Illumina NovaSeq 6000 platform with 100-bp paired-end reads at the Translational Genomics Research Institute (TGen).