in vitro culture for 72 hours with anti CD3/28, Th9-promoting cytokines/antibodies, and retinoic acid
chip antibody
AbCam ab4729
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Cells were chemically cross linked by 1% formaldehyde. Cell lysates were made by appropriate sonication and protein-DNA complexes were isolated with antibody. Aliquots of genomic DNA (input) and immunoprecipitated samples were treated with proteinase K, heated to induce de-cross linking, and purified using columns (D4014, Zymo). After recovering purified DNA, 5ng or more of DNA was used to generate libraries according to the vendor's manual for the Illumina platform (Cat#0344; NuGEN). The purified libraries were multiplexed and captured on an Illumina flow cell for cluster generation. Libraries were sequenced for 50 single read cycles on HiSeq 2500 following the manufacturer's protocols.