characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2014-12-31T17:01:26Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-08T16:52:28Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA2234724
characteristics.characteristic: External Id
characteristics.text: National Research Center for Epigenome Reprogramming Network
characteristics.characteristic: INSDC center name
characteristics.text: 2014-12-31T17:01:26Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-08T16:52:28Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS363976
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-2030:Control sample - undifferentiated 1
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: E14
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0007075
characteristics.characteristic: cell line
characteristics.text: embryonic stem cell
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/CL_0002322
characteristics.characteristic: cell type
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: We purchased normal mouse embryonic stem cells (E14) from the Mutant Mouse Regional Resource Centers (MMRRC). We cultured stem cells in DMEM containing 15% (v/v) FBS (Hyclone), 2 mM L-glutamine, 1% (v/v) nonessential amino acids, 55uM-mercaptoethanol, antibiotics (all from Gibco) and 1,000 U ml-1of ESGRO (Chemicon) on gelatin-coated plates. ESC differentiation was induced by LIF withdrawal of ESC medium in monolayer cultures. PLKO-puro constructs expressing shRNAs were purchased from Sigma. The shRNA constructs targeting Ctbp2 are TRCN0000109335 and TRCN0000307495. Lentiviruses were produced with 293FT cell by co-transfection of 1 mg each of pMD2.G, pMDLg/pRRE, pRSV-rev, pLKO-shRNA using Lipofectamin (Invitrogen). 48 h after transfection, virus-containing medium was collected and filtered by 0.45 mm filters. Polybrene (10 mg ml-1) was added just before infection to target cells and infection was performed for 5 h. Puromysin selection (2 mg ml-1) was performed 48 h after infection for 2 days. As a control, we used pLKO.1 puro vector. RNA was isolated by Trizol reagent (Invitrogen). Sequencing library was prepared using Truseq RNA Sample Preparation kit v2 (IIIumina, CA, USA) according the manufacture's protocols. Briefly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads, fragmented, and converted into cDNAs. Then, adapters were ligated and the fragments were amplified on a PCR.
characteristics.characteristic: description
characteristics.text: undifferentiated
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0000621
characteristics.characteristic: developmental stage
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_10090
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: fresh specimen
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/OBI_0000971
characteristics.characteristic: specimen with known storage state
characteristics.text: 129P2/OlaHsd
characteristics.characteristic: strain
characteristics.text: Control sample - undifferentiated 1
characteristics.characteristic: title