GSM5964811: JNKi_IgG (ChIP-seq); Mus musculus; ChIP-Seq
Cells were washed 2x with ice cold PBS and crosslinked for 10 minutes at room temperature in 1% formaldehyde PBS (ThermoFisher #28906). The cross-linked chromatin was sheared by sonication, providing fragments of 200 - 1000 base pairs in length. Protein-DNA complexes were selectively immunoprecipitated using CREB,c-Jun and igG antibodies. The immunoprecipitated complexes were washed to remove non-specifically bound chromatin, the protein-DNA cross-link was reversed and proteins were removed by proteinase K digest. At least three IPs were pooled for each sample of the three biological replicates, in order to obtain enough DNA precipitated. The DNA associated with the complex was furthermore purified and used for ChIP-seq. The library generation was carried out by the VBCF NGS facility (https://www.viennabiocenter.org/vbcf/next-generation-sequencing/). The chromatin quality was analyzed on an Agilent 2100 Bioanalyzer and a size selection of 200-1000bp was carried out.