Comment[ArrayExpressAccession]	E-GEOD-43658
MAGE-TAB Version	1.1																
Public Release Date	2013-04-01
Investigation Title	Transcriptional co-factor TBLR1 controls lipid mobilization in white adipose tissue																
Comment[Submitted Name]	Transcriptional co-factor TBLR1 controls lipid mobilization in white adipose tissue
Experiment Description	Lipid mobilization (lipolysis) in white adipose tissue (WAT) critically controls lipid turnover and adiposity in humans. While the acute regulation of lipolysis has been studied in detail, the transcriptional determinants of WAT lipolytic activity remain still largely unexplored.  Here we show that the genetic inactivation of transcriptional co-factor transducin beta-like-related (TBLR) 1 blunts the lipolytic response of white adipocytes through the impairment of cAMP-dependent signal transduction. Indeed, mice lacking TBLR1 in adipocytes are defective in fasting-induced lipid mobilization and when placed on a high fat diet show aggravated adiposity, glucose intolerance and insulin resistance. TBLR1 levels are found to increase under lipolytic conditions in WAT of both human patients and mice, correlating with serum free fatty acids (FFA). As a critical regulator of WAT cAMP signaling and lipid mobilization, proper activity of TBLR1 in adipocytes may thus represent a critical molecular checkpoint for the prevention of metabolic dysfunction in subjects with obesity-related disorders. We used microarrays to identify global gene expression in 3T3-L1 adipocytes lacking TBLR1 and compared gene expression to control shRNA treated cells in both basal and isoproterenol stimulated states. We analyzed 12 RNA samples extracted from 3T3-L1 adipocytes that were treated with either control or TBLR1 specific shRNAs and with or without 10 µM isoproterenol for 3 hrs. Three replicates of each condition.																
Term Source Name	ArrayExpress	EFO
Term Source File	http://www.ebi.ac.uk/arrayexpress/	http://www.ebi.ac.uk/efo/efo.owl															
Person Last Name	Sticht	Rohm	Sommerfeld	Strzoda	Jones	Sijmonsma	Rudofsky	Wolfrum	Sticht	Gretz	Zeyda	Leitner	Nawroth	Stulnig	Berriel Diaz	Vegiopoulos	Herzig
Person First Name	Carsten	Maria	Anke	Daniela	Allan	Tjeerd	Gottfried	Christian	Carsten	Norbert	Maximilian	Lukas	Peter	Thomas	Mauricio	Alexandros	Stephan
Person Email	geo@ncbi.nlm.nih.gov																
Person Affiliation	University Heidelberg																
Person Address	ZMF, University Heidelberg, Theodor-Kutzer-Ufer, Mannheim, Germany																
Person Roles	submitter																
Protocol Name	P-GSE43658-1	P-GSE43658-6	P-GSE43658-3	P-GSE43658-8	P-GSE43658-7	P-GSE43658-2	P-GSE43658-4	P-GSE43658-5									
Protocol Description	ID_REF =  VALUE = quantile normalized signal	Hybridization (16h x 45°C) was processed according to the standard Affymetrix protocol.	3T3-L1 preadipocytes were cultured in low (1g/l) glucose DMEM medium containing 10% fetal calf serum (FCS) and 1% penicillin/streptomycin (P/S) and differentiated into mature white adipocytes two days post confluency by the addition of 1 µg/ml insulin, 0.5 mM 3-isobutyl-1-methylxanthine (IBMX), 0.25 µM dexamethasone and 1/1000 volume ABP (50 mg/ml L-ascorbate, 1 mM biotin, 17 mM pantothenate) in high (4.5 g/l) glucose DMEM containing 10% FCS and 1% P/S	The data were analyzed with a commercial software called JMP Genomics, version 4, from SAS. Gene expression profiling was performed using arrays of Mouse430_2 -type from Affymetrix. A Custom CDF Version 13 with Entrez based gene definitions was used to annotate the arrays. The Raw fluorescence intensity values were normalized applying quantile normalization.	Affymetrix GeneArray Scanner3000	Mature adipocytes 9 days after induction of differentiation were transduced with adenoviruses carrying shRNA against TBLR1 or a control sequence at a multiplicity of infection (MOI) of 500. After 3 days, cells were preincubated in Krebs-Ringer buffer for 2 hrs, then cells were stimulated with 10 µM isoproterenol for 3 hrs.	RNeasy Mini Kit (Quiagen)	Biotinylated cRNA were prepared according to the standard Affymetrix protocol.									
Protocol Type	bioassay_data_transformation	hybridization	grow	feature_extraction	image_aquisition	specified_biomaterial_action	nucleic_acid_extraction	labeling									
Experimental Factor Name	TREATMENT	VECTOR															
Experimental Factor Type	treatment	vector															
Comment[SecondaryAccession]	GSE43658																
Comment[GEOReleaseDate]	2013-04-01																
Comment[ArrayExpressSubmissionDate]	2013-01-22																
Comment[GEOLastUpdateDate]	2013-04-01																
Comment[AEExperimentType]	transcription profiling by array																
SDRF File	E-GEOD-43658.sdrf.txt