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Stable conditional expression and effect of C/EBPβ-LIP in adipocytes using the pSLIK system

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Item Type:Article
Title:Stable conditional expression and effect of C/EBPβ-LIP in adipocytes using the pSLIK system
Creators Name:Esteves, C.L. and Kelly, V. and Bégay, V. and Lillico, S.G. and Leutz, A. and Seckl, J.R. and Chapman, K.
Abstract:Murine 3T3-L1 adipocytes are widely used as a cellular model of obesity. However, whereas transfection of 3T3-L1 preadipocytes is straightforward, ectopic gene expression in mature 3T3-L1 adipocytes has proved challenging. Here, we used the pSLIK vector system to generate stable doxycycline-inducible expression of the liver-enriched inhibitor protein isoform of CCAAT/enhancer binding protein (C/EBP) {beta} (C/EBP{beta}-LIP) in fully differentiated 3T3-L1 adipocytes. Because overexpression of C/EBP{beta}-LIP impairs adipocyte differentiation, the C/EBP{beta}-LIP construct was first integrated in 3T3-L1 preadipocytes but expression was induced only when adipocytes were fully differentiated. Increased C/EBP{beta}-LIP in mature adipocytes down-regulated C/EBP{beta} target genes including 11{beta}-hydroxysteroid dehydrogenase type 1, phosphoenolpyruvate carboxykinase and fatty acid binding protein 4, but had no effect on asparagine synthetase, demonstrating that transcriptional down-regulation by C/EBP{beta}-LIP in 3T3-L1 adipocytes is not a general effect. Importantly, these genes were modulated in a similar manner in adipose tissue of mice with genetically increased C/EBP{beta}-LIP levels. The use of the pSLIK system to conditionally express transgenes in 3T3-L1 cells could be a valuable tool to dissect adipocyte physiology.
Keywords:Adipocyte, Stable Transfection, pSLIK, C/EBP{beta}-LIP, Animals, Mice
Source:Journal of Molecular Endocrinology
Page Range:91-98
Date:4 June 2013
Official Publication:https://doi.org/10.1530/JME-13-0029
PubMed:View item in PubMed

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