leukemic cell line with an 8;21 chromosome translocation
passages
4-5 passages
chip antibody
HA-Tag (C29F4) Rabbit mAb (CST #3724)
genotype/variation
overexpress HEB
cell line
Kasumi-1
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
At 4 days after retrovirus infection of MIGR1 and E-protein, GFP+ Kasumi-1 cells were harvested by flow sorting and RNA was extracted with TRIzol Reagent (Invitrogen). (RNA-seq) At 3 days after retrovirus infection of E-protein, GFP+ Kasumi-1 cells were sorted by BD FACSARIA III through FITC channel. Then they were crosslinked with 1% formaldehyde for 10 mins and then stopped by 125 mM Glycine. After lysis and sonication, 3 μg of HA-Tag (C29F4) Rabbit mAb (CST #3724) was incubated with chromatin sample overnight at 4 ℃, and 20μl protein A/G beads (Smart lifesciences SA032005) were used for immunoprecipitation. Finally, DNA were purified with PCR recovery kit (QIAGEN #28006).(ChIP-seq) Libraries were generated using the TruSeq RNA Sample Preparation Kit (Illumina #RS-122-2001) and sequenced with Illumina HiSeq2500.(RNA-seq) Library preparation was according to the manufacture's guidelines (KAPA Biosystems KK8503) and sequencing procedures were carried out according to Illumina protocols with minor modifications (Illumina, San Diego).(ChIP-seq)