title: Methylation mimic mutations of progesterone receptor AF1 impair gene-specific regulation through stabilized chromatin interactions
library_construction_protocol: After treatment, cells were crosslinked with formaldehyde at a final concentration of 1% for 10 minutes at room temperature. The cross-linking was quenched by the addition of glycine to a final concentration of 0.125 M. Cells were then harvested, lysed, and subjected to sonication using Diagenode Bioruptor Pico.  Chromatin immunoprecipitation was performed using anti-PR antibody H-190 (Santa Cruz Biotechnology Inc., Dallas, TX, USA, LOT number sc-7208). The lysate was incubated with 5 ug anti-PR antibody overnight at 4 °C. Protein-chromatin complex was captured using Pierce™ ChIP-grade Protein A/G Magnetic Beads (ThermoFisher Scientific, Cat. #26162). Chromatin was eluted, and crosslinks were reversed by treatment with RNase A and Proteinase K. Input samples and ChIP'ed DNA was purified using QIAquick PCR Purification Kit (Qiagen). DNA concentrations were quantified using Qubit Fluorometer (ThermoFisher Scientific). The samples were submitted to Genome Institute of Singapore (Agency for Science, Technology and Research) for library preparation and paired-end sequencing using Illumina HiSeq4000 platform.