ChIP experiments were performed as previously described (Schmidt D, Wilson MD, Spyrou C, Brown GD, Hadfield J, Odom DT. ChIP-seq: using highthroughput sequencing to discover protein-DNA interactions. Methods. Jul 2009;48(3):240-248). Approximately 5 x 107 cells were used for each IP. ChIP-Seq experiments were conducted on two replicates of K1ER cells induced with 200 nM Tamoxifen for 2 hrs, and two replicates of HUDEP-2 cells. The antibodies used were α-KLF1 (PIEPA5-18031, Thermo Scientific), anti-goat IgG (SCZSC-2028, Santa Cruz) as a negative control. Library preparation was performed using the TruSeq DNA Sample Preparation Kit (FC-121-2001, Illumina) according to the manufacturer’s instructions with minor modifications. Adapter sequences were diluted 1/40 before use and following adapter ligation, the library size extracted from the gel was 100-280 bp (excluding adapters) in line with the size of sonicated fragments. Libraries (2 inputs and 2 IP samples) were multiplexed into 2 lanes such that there were 2 samples per lane. Samples were sequenced using 50 bp single read chemistry on the HiSeq 2500 (Illumina).