Identification of Smad2 and Smad3 specific binding sites in a breast cancer cell line model

We identify Smad2 and Smad3 specific binding site in a breast cancer breast cell model MDA-MB-231 using LAP-tag GFP fusion proteins under the control of endogenous regulatory elements. Stable cell lines were generated via transfection with a BAC followed by antibiotics selection. We show that LAP-fused SMad2/3 can be used to recapitulate native TGFB signaling. Smad2/Smad3 specific binding sites were identified by ChIP-seq against GFP before and after TGFB treatment. Genomic DNA was sequenced via single end sequencing on Illumina HiSeq Chromatin immunoprecipitation against GFP in fusion protein before and after TGFB treatment