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

Esteves, C.L.; Kelly, V.; Bégay, V.; Lillico, S.G.; Leutz, A.; Seckl, J.R.; Chapman, K.

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., Kelly, V., Bégay, V., Lillico, S.G., Leutz, A., 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
ISSN:	0952-5041
Publisher:	BioScientifica
Volume:	51
Number:	1
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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