characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2017-05-22T17:14:24Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2016-02-01T22:20:11Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA3862445
characteristics.characteristic: External Id
characteristics.text: INSERM U1170, Equipe Labellisee Ligue Contre le Cancer, Villejuif, France Institut Gustave Roussy, Villejuif, France Universite Paris Diderot, Paris, France
characteristics.characteristic: INSDC center name
characteristics.text: 2017-05-22T17:14:24Z
characteristics.characteristic: INSDC first public
characteristics.text: 2016-02-01T22:20:11Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS1049579
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-4367:AMKL7 ETO2
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: Human patient's cells from xenograft
characteristics.characteristic: cell type
characteristics.text: human
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: common name
characteristics.text: Protocols: The human MO7e and MO7e-KI AMKL cell lines were cultured in MEM- supplemented with 20% FBS, Penicillin (100U/mL)-Streptomycin (100g/mL) and 2mM L-Glutamine (Gibco), and 5 ng/ml of human GM-CSF (PeproTech). HEL cells are cultured in RPMI1640 supplemented with 10% FBS, Penicillin (100U/mL)-Streptomycin (100g/mL), 2mM L-Glutamine (Gibco). AMKL cells obtained directly from immunodeficient animals or after purification by flow cytometry were cultured in RPMI 1640 medium supplemented with 20% FBS, Penicillin (100U/mL)-Streptomycin (100g/mL) and 2mM L-Glutamine (Gibco), and 10 ng/ml each of human SCF, IL3, IL6, IL11, TPO, 10U/ml EPO and 5 ng/ml of GM-CSF (PeproTech). ChIP protocol was adapted from the MagnaChIP kit protocol (Millipore). Cells (AMKL and MO7e-KI) were fixed with 1% formaldehyde, lysed with Cell Lysis and then Nuclear Lysis buffers respecting concentration of 20.106 cells per mL, and finally sonicated (30-min cycle on Covaris apparatus; KBioscience). Sheared chromatin was immunoprecipitated overnight using the following antibodies: anti-CBFA2T3 (Abcam, ab33072), anti-ERG (Santa Cruz ; SC354), anti-GATA3 (Santa Cruz ; mix 50/50 of SC268X and SC269X), anti-H3K27ac (ActiveMotif, #39133), anti-H3K4me1 (Diagenode ; #C15410037), anti-H3K4me3 (Diagenode, #C15410003), anti-H3K27me3 (Cell Signaling, #9733S) and rabbit IgG (Upstate, #12-370). 1/10 of the sheared chromatin was used as a reference (Input). Immune complex collection was realized with Protein G Sepharose (Sigma-Aldrich; P3296), 1h30 at +4C. For GFP-ChIP, GFP-Trap agarose beads were used (Chromotek). Rinses were done according to MagnaChIP kit protocol with Low salt, High salt and LiCL immune complex wash buffers. Finally, elution was realized according the IPure Kit protocol (Diagenode, Cat No C03010012) following manufacturers instructions. Enriched DNA from ChIP and Input DNA fragments were end-repaired, extended with an 'A' base on the 3end, ligated with indexed paired-end adaptors (NEXTflex, Bioo Scientific) using the Bravo Platform (Agilent), size-selected after 4 cycles of PCR with AMPure XP beads (Beckman Coulter) and amplified by PCR for 10 cycles more.
characteristics.characteristic: description
characteristics.text: acute megakaryoblastic leukaemia
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_1001932
characteristics.characteristic: disease
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: AMKL7 ETO2
characteristics.characteristic: title