Mus musculus
Protocols: For generation of Meer cells primary hematopoietic precursor cells were isolated from the bone marrow of Meer mice that carry a knock-in of an ENL-ER (mutant estrogen receptor ligand binding domain) portion into the mouse Mll locus. (Lit: Takacova, S. et al. Cancer Cell 21, 517-31, 2012). CD117 (c-kit) positive cells were isolated by magnetic enrichment according to the instructions of the manufacturer (CD117 enrichment kit, Miltenyi Biotech, Bergisch-Gladbach, Germany) and initially plated in methocel (MethoCult M3234, StemCellTechnologies, Koln, Germany) to purge residual stroma cells that cannot grow in semisolid media. After two rounds of growth in methylcellulose cells were expanded in RPMI1640 supplemented with 10% FCS. All media contained 50ng/ml recombinant murine (rm) SCF, 10ng/ml of rm-IL3, IL6, GM-CSF (cytokines were bought from Miltenyi Biotech, Bergisch-Gladbach, Germany) and for activation of the estrogen receptor 100nM 4-hydroxytamoxifen (TAM, Sigma, Taufkirchen, Germany). ChIP was done exactly as described in (Milne, T.A., Zhao, K. & Hess, J.L. Chromatin immunoprecipitation (ChIP) for analysis of histone modifications and chromatin-associated proteins. Methods Mol Biol 538, 409-23 (2009) with the exception that protein G-coupled paramagnetic beads from CellSignalingTechnologies (CST/NEB, Frankfurt, Germany) were used as precipitating agent. Antibodies and concentrations used : anti-H2AUb, Cell Signaling Technologies (#8240) 10µl; anti-H2BUb, Cell Signaling Technologies (#5546) 5µl ChIP Seq library construction with Illumina Kit according to the instructions of the manufacturer
ENA-FIRST-PUBLIC
2018-08-20T17:02:52Z
ENA-LAST-UPDATE
2017-03-10T14:51:49Z
External Id
SAMEA103906300
INSDC center name
Friedrich Alexander University Erlangen-Nurnberg FAU Department Biology
INSDC first public
2018-08-20T17:02:52Z
INSDC last update
2017-03-10T14:51:49Z
INSDC status
public
Submitter Id
E-MTAB-5569:H2AUb_smpl1
broker name
ArrayExpress
cell line
Meer
cell type
hematopoietic precursor cell
common name
house mouse
genotype
Mll-ENL-ER
sample name
E-MTAB-5569:H2AUb_smpl1
scientific_name
Mus musculus