Stable conditional expression and effect of C/ebpβ-LIP in adipocytes using the pSLIK system
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 β (C/ebpβ (Cebpb)) (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.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2013 |
|---|---|
| Erschienen: |
2013 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:51 |
|---|---|
| Enthalten in: |
Journal of molecular endocrinology - 51(2013), 1 vom: 14., Seite 91-8 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Esteves, Cristina L [VerfasserIn] |
|---|
| Links: |
|---|
| Anmerkungen: |
Date Completed 07.01.2014 Date Revised 21.10.2021 published: Electronic-Print Citation Status MEDLINE |
|---|
| doi: |
10.1530/JME-13-0029 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM227004000 |
|---|
| LEADER | 01000naa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM227004000 | ||
| 003 | DE-627 | ||
| 005 | 20231224072706.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231224s2013 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1530/JME-13-0029 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n0756.xml |
| 035 | |a (DE-627)NLM227004000 | ||
| 035 | |a (NLM)23620165 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Esteves, Cristina L |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Stable conditional expression and effect of C/ebpβ-LIP in adipocytes using the pSLIK system |
| 264 | 1 | |c 2013 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 07.01.2014 | ||
| 500 | |a Date Revised 21.10.2021 | ||
| 500 | |a published: Electronic-Print | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a 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 β (C/ebpβ (Cebpb)) (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 | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a C/EBPβ-LIP | |
| 650 | 4 | |a adipocyte | |
| 650 | 4 | |a pSLIK | |
| 650 | 4 | |a stable transfection | |
| 650 | 7 | |a CCAAT-Enhancer-Binding Protein-beta |2 NLM | |
| 650 | 7 | |a Fabp4 protein, mouse |2 NLM | |
| 650 | 7 | |a Fatty Acid-Binding Proteins |2 NLM | |
| 650 | 7 | |a 11-beta-Hydroxysteroid Dehydrogenase Type 1 |2 NLM | |
| 650 | 7 | |a EC 1.1.1.146 |2 NLM | |
| 650 | 7 | |a Phosphoenolpyruvate Carboxykinase (GTP) |2 NLM | |
| 650 | 7 | |a EC 4.1.1.32 |2 NLM | |
| 700 | 1 | |a Kelly, Val |e verfasserin |4 aut | |
| 700 | 1 | |a Bégay, Valérie |e verfasserin |4 aut | |
| 700 | 1 | |a Lillico, Simon G |e verfasserin |4 aut | |
| 700 | 1 | |a Leutz, Achim |e verfasserin |4 aut | |
| 700 | 1 | |a Seckl, Jonathan R |e verfasserin |4 aut | |
| 700 | 1 | |a Chapman, Karen E |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Journal of molecular endocrinology |d 1996 |g 51(2013), 1 vom: 14., Seite 91-8 |w (DE-627)NLM012626864 |x 1479-6813 |7 nnns |
| 773 | 1 | 8 | |g volume:51 |g year:2013 |g number:1 |g day:14 |g pages:91-8 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1530/JME-13-0029 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 51 |j 2013 |e 1 |b 14 |h 91-8 | ||