title: NextSeq 500 sequencing; Herbal formula YYJD induces lung cancer cell apoptosis via activation of EGR1
design: Herbal formula YYJD induces lung cancer cell apoptosis via activation of EGR1
library_construction_protocol: A549 was maintained in RPMI 1640 medium (Corning) supplemented with 10% FBS (Gibco) and 100 units per ml penicillin–streptomycin solution at 37℃, 5% CO2 in a humidified incubator, then treated with anti-tumor drug YYJD.  Cells were covalently cross-linked by incubating cells in 1% formaldehyde (Sigma-Aldrich) for 10 minutes at room temperature and incubated with 125 mM glycine for 5 minutes to quench cross-linking reaction. The fragmented chromatin fragments were immunoprecipitated with Protein A+G Magnetic beads (Millipore) coupled with anti-EGR1 antibody (4154, Cell Signaling Technology).  Immunoprecipitated DNAs and input DNAs were end-repaired and ligated adapters to the DNA fragments using NEBNext Ultra End-Repair/dA-Tailing Module (E7442, NEB) and NEBNext Ultra Ligation Module (E7445, NEB).
attributes.tag: Experimental Factor: compound
attributes.value: YYJD
attributes.tag: Experimental Factor: immunoprecipitate
attributes.value: anti-EGR1