GSM5381234: Treated 1; Mus musculus; ChIP-Seq
ChIP-seq analysis of COUP-TFII (NR2F2) and HNF4α  following 2 hrs TCDD exposure was performed using the FactorPath services by Active Motif (Carlsbad, CA) on ~100mg frozen liver sections. Livers cut in small pieces were incubated in 1% formaldehyde for 15 min at room temperature. Cross-linking was halted by the addition of 0.125 M glycine and tissue was homogenized by Dounce homogenizer followed by shearing DNA by sonication to achieve an average length of 300 – 500 bp. A total of 30 μg of precleared sheared chromatin with protein A agarose beads was immunoprecipitated using 5 μL of rabbit anti-NR2F2 polyclonal IgG (sc-8987; Santa Cruz, Dallas, TX) or 20uL of rabbit anti-HNF4a polyclonal IgG (61213; Active Motif) overnight at 4°C. Chromatin was eluted from the beads with SDS buffer and cross-links reversed by incubation with RNase and Proteinase K followed by an overnight incubation at 65°C. Libraries for sequencing were prepared by Active Motif and sequenced using NextSeq 500 (1 x 75bp).