title: ATAC-seq from mouse embryonic stem cell and their derivative neural stem cells
design: ATAC-seq from mouse embryonic stem cell and their derivative neural stem cells
library_construction_protocol: Cells were detached with Accutase, counted and 100,000 cells were immediately used for the ATAC-seq experiments. ES (2i/LIF) cells were grown on gelatin-coated culture plastic in embryonic stem cells medium in the presence of MEK inhibitor (PD0325901) and GSK3 inhibition (CHIR99021) with LIF to maintain ESCs in a naive ground state. Cells were split at 80% confluence with a density of 30000 cells/sq.cm. The medium was exchanged daily.  NS cells were grown on laminin-coated plates in neural differentiation medium containing N2 and B27 supplements along with recombinant murine EGF and bFGF to a final concentration of 10 ng/ml. Cells were split at 80% confluence with a density of 35000 cells/sq.cm. The medium was exchanged daily.  CTCF depletion was carried out in CTCF-AID-GFP ES and NS Cells by treating with 500uM of indole acetic acid (IAA) dissolved in culture media for 24h ATAC-seq procedure was performed with ATAC-Seq kit (Active Motif, 53150), according to manufacturer's instructions.