characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2017-09-21T17:01:36Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2017-09-11T11:24:19Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA104289171
characteristics.characteristic: External Id
characteristics.text: SV-IBI-EPFL
characteristics.characteristic: INSDC center name
characteristics.text: 2017-09-21T17:01:36Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-09-11T11:24:19Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS1914149
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-5159:IBA_Day0_NFI
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: immortalised brown preadipocyte
characteristics.characteristic: cell line
characteristics.text: brown preadipocyte
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/CL_0002334
characteristics.characteristic: cell type
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: Murine derived brown pre-adipocytes (IBA) were cultured in high-glucose Dulbecco’s modified Eagle’s medium (DMEM, Life Technologies) supplemented with 10% fetal calf serum (FCS, AMIMED), with 100x penicillin/streptomycin/glutamine (100x Pen/Strep/Glutamine, Life Technologies) in a 5% CO2 humidified atmosphere at 37C and maintained at less than 80% confluence before passaging Differentiation of IBAs was induced by exposing confluent cells (Day 0) to DMEM containing 10% FCS supplemented with 0.5 mM 3-isobutyl-1-methylxanthine, 1 uM dexamethasone, 1.96 nM insulin (Sigma, stock solution of 167uM prepared in PBS), 125 uM indomethacin (Sigma, stock solution of 18mM prepared in absolute ethanol), 1nM T3 (3,3’,5-Triiodo-L-thyronine sodium salt, Sigma, prepared in 45mM KOH). After exactly 28 hours, the induction medium was replaced with adipocyte differentiation medium consisting of 1nM T3 and 1.96 nM insulin Murine brown pre-adipocytes were collected at Day 0, 2 hours, Day 1, Day 2 and Day 4. The cells were fixed as described previously (Raghav and Deplancke, 2012) and stored at -80C. Ten million cells were used for each IP. The ChIP experiment was performed as using magnetic beads protocol (Dynabeads) (H3K27ac and NFI). Multiplexed libraries were prepared using barcoded adapters for each sample following the protocol described in (Raghav and Deplancke, 2012) with slight modifications. In brief, ChIP-DNA fragments (5-10ng) were end-repaired using an End-IT DNA end repair kit (Epicentre Technologies) followed by addition of an A-base and ligation of bar-coded adapters. After ligation incubation, DNA was cleaned up using AMPure XP beads (Agencourt). Two biological replicates for five time points (Day 0, 2h, Day 1, Day 2 and Day 4) were prepared for probing H3K27ac levels during differentiation, while one biological replicate at Day 0 and Day 4 was prepared for NFI.
characteristics.characteristic: description
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: brown preadipocyte
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/CL_0002334
characteristics.characteristic: progenitor cell type
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: IBA_Day0_NFI
characteristics.characteristic: title