GSM5900089: CRE_IKZF-WT_H3K27ac_rep2; Mus musculus; ChIP-Seq
B-ALL cells were counted and fixed with 1% formaldehyde for 10 min at room temperature and quenched by 125 mmol l-1 glycine. B-ALL cells were spiked with fixed Drosophila S2 cells, and subjected to nuclear extraction. Nuclei were lysed with SDS lysis buffer and sonicated by Bioruptor (Diagenode). After sonication, cell debris was removed by centrifugation (13,000 rpm, 10 min), while supernatant was diluted and pre-cleared by incubation with a blend of isotype control IgG, Dynabeads Protein A and Dynabeads Protein G (Invitrogen). ChIP was performed using antibodies specifically recognizing either H3K27ac (Active motif, #39133) or H3K4me3 (Millipore, MC315). ChIPed DNA was reverse-crosslinked overnight at 65℃, and purified by QIAquick PCR purification kit (Qiagen). For ChIP-seq of IKZF1 and IKZF3, B-ALL cells were crosslinked by 2 mmol l-1 disuccinimidyl glutarate for 45 min and 1% formaldehyde for 10 min at room temperature before chromatin enrichment and library construction. These antibodies were used for ChIP-seq of  IKZF1 (GeneTex, GTX129438) and IKZF3 (CST, D1C1E). β-catenin ChIP was performed by Active Motif, Inc. (Carlsbad, CA, USA) using the monoclonal antibody CAT-15 (Thermo). ChIP-seq libraries were constructed by a SMARTer ThruPLEX DNA-seq Kit (Takara).