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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 |
| 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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