4C-seq analysis in TAL1-FKBP12 Jurkat cells

4C analysis was performed in TAL1-FKBP12 Jurkat cells to analyze chromatin-chromatin interactions after DMSO or dTAG-13 treatment. Restriction enzyme digestion was performed with N1aIII (NEB) followed by overnight ligation and subsequent reverse crosslinking. DNA fragments were purified to yield the 3C template. 3C template was then subjected to a second restriction enzyme digestion by DpnII (NEB), overnight ligation and sample purification were repeated to yield the 4C template. Library preparation involved two PCR steps to amplify the fragments ligated to the sequence of interest followed by a second PCR to incorporate sequencing adapters.