GSM8880286: OVSAHO, IFNγ Treatment, STAT3 (Input); Homo sapiens; ChIP-Seq
Cells were crosslinked using 1% formaldehyde (ThermoFisher, Waltham, MA) per 10 mL cell culture medium and placed on a rocker to mix for exactly 10 minutes at room temperature. Glycine (Fisher Scientific, Hampton, NH) was then added to a final concentration of 125 mM to each plate and mixed well by slowly swirling 10 times and incubated at room temperature for 5 minutes. Fixed cells were washed with cold 1X TBS (Bio Rad, Hercules, CA) and scraped into a 15 mL collection tube, centrifuged at 2500 rpm for 5 minutes at 4°C, and supernatant was removed. Another 3 mL of cold 1X TBS was added to each tube to wash, spun again at 2500 rpm for 5 minutes at 4°C, and supernatant was removed. For each treatment ChIP was performed with STAT1 (#SC417, Santa Cruz Biotechnology, Dallas, TX) and STAT3 (#CST9139, Cell Signaling, Danvers, MA) antibodies Sequencing libraries were prepared using the Ribicon Genomics ThruPLEX DNA-seq kit (Takara Bio, Mountain View, CA) and sequencing was performed by the Mayo Clinic Genome Analysis Core using an Illumina HiSeq 4000.