ChIP-Atlas: ERX075981

ERX075981
Genome-wide Analysis of Sp2 Occupancy and Expression Profiling Establish Sp2 as a Sequence-specific Transcription Factor Regulating Fundamental Cellular Processes - ChIPseq

library_name: MEF deltaSp2 antiSp2-AB1
library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: MEFs were maintained at 37 degrees celcius in 50 % DMEM High Glucose and 50 % Ham's F10 Medium (PAA) supplemented with 10% fetal bovine serum (PAA) and 1X Penicillin and Streptomycin (PAA). ChIP-qPCR experiments were performed essentially as described previously (Stielow et al., 2008, see below) using the OneDay ChIP kit (Diagenode) according to the manufacturer's instructions.<br> Briefly, MEFs and HEK293 cells were incubated with 1% formaldehyde for 10 min at room temperature followed by quenching with 150 mM glycine for 5 min. After sonification, 100 microgram of chromatin was used for single immunoprecipitations using 2 microgram of self-made Sp2 antibody (Baur et al., 2010). Ten immunoprecipitations were pooled for ChIP-seq and purified by the QIAquick PCR Purification kit (Qiagen). 10 ng of inmunoprecipitated DNA fragments were subjected to paired-end sequencing on an Illumina IIx Genome Ananlyzer.<br><br>Stielow, B., et al. (2008). SUMO-modified Sp3 represses transcription by provoking local heterochromatic gene silencing. EMBO Rep 9, 899-906.