Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of mouse R1 mESC exposed to butyrate

ABSTRACT: To assess the effect of sodium butyrate exposure on mouse ESC grown without self-renewal support (I.e. without feeders or LIF) - these cells were compared to cells grown without sodium buytrate, but with added mouse LIF via Agilent whole mouse genome array Experiment Overall Design: R1 mESC were grown under 2 conditions, with mLIF on gelatin or without LIF on gelatin for 15 passages to compare change in the RNA profile

ORGANISM(S): Mus musculus

SUBMITTER: Brig Mecham

PROVIDER: E-GEOD-15107 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:This experiment is part of the FunGenES project (FunGenES - Functional Genomics in Embryonic Stem Cells partially funded by the 6th Framework Programme of the European Union, http://www.fungenes.org).The experiment was conducted at the Unyversity of Bath, Bath, UK. Aim is to identify genes regulated by PI3K-dependent signaling in undifferentiated ES cells. Materials and methods: E14tg2a cell line was used. Cells were cultured in Knock-out DMEM supplemented with 15% (v/v) Knock-out serum replacement, 1000U/ml LIF, 2mM glutamine, 50 ?M 2-mercaptothanol and 1x NEAA (as described in Handbook of work methods, 4.3.1.1). Cells were plated at 5x105 per 10cm gelatin-coated TC dish for 24h. ES cells were then starved for 24h in media lacking LIF and pre-incubated for 30 minutes with 5 <B5>M LY294002 (the concentration of this PI3K inhibitor that we have shown to perturb self-renewal in the presence of LIF and which knocks down PI3K signals) or DMSO alone. Cells were then stimulated with 103U/ml mLIF (Chemicon) for 2h (which gave maximal activation of STAT3), 24h, 48h or 72h.</p>\n<h3>Relationships between samples</h3>\n <p ALIGN="JUSTIFY">Samples were either maintained in LIF or LIF plus LY294002 (PI3K inhibitor) for the required treatment times.</p>\n<h3>Treatments</h3>\n <p ALIGN="JUSTIFY">Treatment times with PI3K inhibitor LY294002 at 5 <B5>M <96> 2h (n=5), 24h (n=3), 48h (n=3), 72h (n=3). DMSO was used as vehicle alone treatment control.</p>\n<h3>Culture conditions</h3>\n <p ALIGN="JUSTIFY">Gelatin-coated TC dishes (Nunc)</p>\n<h3>RNA isolation method</h3>\n <p ALIGN="JUSTIFY">RNA Easy with on-column DNase treatment</p>\n<h3>Experimental description</h3>\n <p ALIGN="JUSTIFY">This information is included above. Briefly, E14tg2a ES cells were either maintained in LIF or LIF plus LY294002 (5 <B5>M) for 2, 24, 48 or 72 hours, after which RNA was extracted from each sample and sent for expression profiling. During this time-course, only 72h time points plus LY294002 showed phenotypic evidence of differentiation. </p>\n

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Transcription profiling of mouse R1 mESC exposed to butyrate

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