ChIP-Seq of SMARCAD1 and KAP1 in male mouse embryonic stem cells (E14)

The ATP dependent chromatin remodeler SMARCAD1 and the co-repressor KAP1 (Trim28/Tif1 beta) interact directly. We wanted to understand the interplay between these two proteins in the context of chromatin; in particular at repetitive sequences. Therefore, we carried out an investigation in E14 ES cells of the genome wide binding behaviour of KAP1 and FLAG-tagged SMARCAD1; both wild-type SMARCAD1 and a catalytically inactive mutant. The E14 cells analysed were either wild-type or depleted for endogenous SMARCAD1 protein. FLAG and KAP1 ChIP was performed on double-cross linked chromatin and sequenced on an Illumina HiSeq1500 with matched input controls. A FLAG antibody precipitation was carried out on cells not expressing FLAG tagged protein as an additional control.