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

Cristina L. Esteves*, Val Kelly, Valerie Begay, Simon G. Lillico, Achim Leutz, Jonathan R. Seckl, Karen E. Chapman

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

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) β (C/EBPβ-LIP) in fully differentiated 3T3-L1 adipocytes. Because overexpression of C/EBPβ-LIP impairs adipocyte differentiation, the C/EBPβ-LIP construct was first integrated in 3T3-L1 preadipocytes but expression was induced only when adipocytes were fully differentiated. Increased C/EBPβ-LIP in mature adipocytes down-regulated C/EBPβ target genes including 11β-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/EBPβ-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/EBPβ-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.
Original languageEnglish
Pages (from-to)91-98
Number of pages8
JournalJournal of molecular endocrinology
Volume51
Issue number1
DOIs
Publication statusPublished - 25 Apr 2013

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