NODEGenomicsCui-Cui LiuChIP-Seqhttps://www.biosino.org/node/experiment/detail/OEX00025490Illumina HiSeq 2000PairedChIP1['cell Line']['Cancer']Briefly, 107 to 108 TNBC cells (Control or EMSY) were harvested, crosslinking was performed with 1% formaldehyde or 1% paraformaldehyde for 10 minutes. To enhance cell lysis, we ran the lysate through a 27g needle three times and flash froze it in −80°C. Sonication was then performed with the Misonix 4000 water bath sonication unit at 15% amplitude for 20 minutes. Protein/DNA complex was precipitated by specific antibodies against H3K4me2/3 (CST) and IgG control (Millipore, Burlington, MA, USA). DNA was purified using DNA Purification Kit (Qiagen, Hilden, Germany). PCR was performed using Phusion High Fidelity PCR master mix. ChIP-enriched chromatins were sequenced on Illumina HiSeq 2000 platforms according to the manufacture’s protocol.NODEfalseOEX_Cui-Cui_2311201516To validate our observation, we performed chromatin immunoprecipitation (ChIP) seq of H3K4me1/2/3 between control and EMSY group, and the result showed that there were high H3K4me2/3 occupancies in the key regulatory promoter regions of stem cell markers, such as NANOG, POU5F1 and SOX2, in EMSY-overexpressing cells.2023-11-202023-11-20OEX00025490OEP00004750