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KLF13 promotes porcine adipocyte differentiation through PPARγ activation.

Jiang S, Wei H, Song T, Yang Y, Zhang F, Zhou Y, Peng J, Jiang S - Cell Biosci (2015)

Bottom Line: Porcine adipocyte differentiation was significantly attenuated by the addition of siRNA against KLF13, whereas overexpression of KLF13 resulted in enhanced porcine adipocyte differentiation.In addition, knockdown and/or overexpression of KLF13 in 3 T3-L1 cells all did not influence expression of PPARγ2.Collectively, our results suggest that KLF13 exist as a key pro-adipogenic transcription factor through transactivating PPARγ expression in porcine adipocyte differentiation, whereas no such effect was detected in mouse adipocyte differentiation.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal Nutrition and Feed Science, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan, P. R. China.

ABSTRACT

Background: Adipogenesis is tightly controlled by a complex network of transcription factors acting at different stages of differentiation. Kruppel-like factors (KLFs) as a family of zinc-finger transcription factors play diverse roles during cell differentiation and development in mammals.

Results: In the present study, we showed that KLF13 acts as a key regulator regulating porcine adipocyte differentiation. The expression of KLF13 was markedly up-regulated during the early stage of porcine adipocyte differentiation, which was followed by expression of PPARγ. Porcine adipocyte differentiation was significantly attenuated by the addition of siRNA against KLF13, whereas overexpression of KLF13 resulted in enhanced porcine adipocyte differentiation. Using promoter deletion and mutation analysis, we identified a KLF13-binding site within -593/-577 region of the porcine PPARγ proximal promoter, indicating that KLF13 directly interacts with porcine PPARγ promoter. However, inhibition of KLF13 by siRNA did not impair mouse adipocyte differentiation. In addition, knockdown and/or overexpression of KLF13 in 3 T3-L1 cells all did not influence expression of PPARγ2.

Conclusions: Collectively, our results suggest that KLF13 exist as a key pro-adipogenic transcription factor through transactivating PPARγ expression in porcine adipocyte differentiation, whereas no such effect was detected in mouse adipocyte differentiation.

No MeSH data available.


Effect of KLF13 on the expression of PPARγ during adipogenic differentiation of porcine ASVC. a Analysis of transcription factor site within the PPARγ promoter. Genomatix MatInspector was used to identify transcription factor site within the promoter of PPARγ from 2000 bp upstream of the translation initiation site. b Effect of KLF13 knockdown on the expression of PPARγ during adipogenic differentiation of porcine ASVC. After 1 days transfection of KLF13 siRNA, SV cells were stimulated in adipogenic induction medium for 3 days. Western-blot was used to determine the protein expression of PPARγ. c Effect of KLF13 overexpression on the expression of PPARγ during adipogenic differentiation of porcine ASVC. After 1 days transfection of pcDNA3.1-KLF13, SV cells were stimulated in adipogenic induction medium for 3 days. Western-blot was used to determine the protein expression of PPARγ. Values are represented as mean ± SD. (n = 3) *P < 0.05
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Fig3: Effect of KLF13 on the expression of PPARγ during adipogenic differentiation of porcine ASVC. a Analysis of transcription factor site within the PPARγ promoter. Genomatix MatInspector was used to identify transcription factor site within the promoter of PPARγ from 2000 bp upstream of the translation initiation site. b Effect of KLF13 knockdown on the expression of PPARγ during adipogenic differentiation of porcine ASVC. After 1 days transfection of KLF13 siRNA, SV cells were stimulated in adipogenic induction medium for 3 days. Western-blot was used to determine the protein expression of PPARγ. c Effect of KLF13 overexpression on the expression of PPARγ during adipogenic differentiation of porcine ASVC. After 1 days transfection of pcDNA3.1-KLF13, SV cells were stimulated in adipogenic induction medium for 3 days. Western-blot was used to determine the protein expression of PPARγ. Values are represented as mean ± SD. (n = 3) *P < 0.05

Mentions: The data shown above suggested that KLF13 would regulate PPARγ and C/EBPα gene expression. To assess whether KLF13 directly activates the PPARγ and C/EBPα gene, we next predicted the binding of KLF13 to the promoters of PPARγ and C/EBPα gene. Bioinformatics analysis showed that only PPARγ gene promoter had a potential binding site (CTCCC) for KLF13. No canonical TATA box was found in the PPARγ promoter region close to the transcription initiation site, whereas a CAAT box was found in the PPARγ promoter (Fig. 3a). Thus, we proposed PPARγ as potential KLF13 direct target during the process of porcine adipocyte differentiation. Real-time PCR analysis has shown that the PPARγ was coordinately regulated by KLF13 in ASVC (Fig. 2c,e). Furthermore, western blot analysis also confirmed that PPARγ expression was coordinately regulated by KLF13 in ASVC under adipogenic induction condition (Fig. 3b,c).Fig. 3


KLF13 promotes porcine adipocyte differentiation through PPARγ activation.

Jiang S, Wei H, Song T, Yang Y, Zhang F, Zhou Y, Peng J, Jiang S - Cell Biosci (2015)

Effect of KLF13 on the expression of PPARγ during adipogenic differentiation of porcine ASVC. a Analysis of transcription factor site within the PPARγ promoter. Genomatix MatInspector was used to identify transcription factor site within the promoter of PPARγ from 2000 bp upstream of the translation initiation site. b Effect of KLF13 knockdown on the expression of PPARγ during adipogenic differentiation of porcine ASVC. After 1 days transfection of KLF13 siRNA, SV cells were stimulated in adipogenic induction medium for 3 days. Western-blot was used to determine the protein expression of PPARγ. c Effect of KLF13 overexpression on the expression of PPARγ during adipogenic differentiation of porcine ASVC. After 1 days transfection of pcDNA3.1-KLF13, SV cells were stimulated in adipogenic induction medium for 3 days. Western-blot was used to determine the protein expression of PPARγ. Values are represented as mean ± SD. (n = 3) *P < 0.05
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Related In: Results  -  Collection

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Fig3: Effect of KLF13 on the expression of PPARγ during adipogenic differentiation of porcine ASVC. a Analysis of transcription factor site within the PPARγ promoter. Genomatix MatInspector was used to identify transcription factor site within the promoter of PPARγ from 2000 bp upstream of the translation initiation site. b Effect of KLF13 knockdown on the expression of PPARγ during adipogenic differentiation of porcine ASVC. After 1 days transfection of KLF13 siRNA, SV cells were stimulated in adipogenic induction medium for 3 days. Western-blot was used to determine the protein expression of PPARγ. c Effect of KLF13 overexpression on the expression of PPARγ during adipogenic differentiation of porcine ASVC. After 1 days transfection of pcDNA3.1-KLF13, SV cells were stimulated in adipogenic induction medium for 3 days. Western-blot was used to determine the protein expression of PPARγ. Values are represented as mean ± SD. (n = 3) *P < 0.05
Mentions: The data shown above suggested that KLF13 would regulate PPARγ and C/EBPα gene expression. To assess whether KLF13 directly activates the PPARγ and C/EBPα gene, we next predicted the binding of KLF13 to the promoters of PPARγ and C/EBPα gene. Bioinformatics analysis showed that only PPARγ gene promoter had a potential binding site (CTCCC) for KLF13. No canonical TATA box was found in the PPARγ promoter region close to the transcription initiation site, whereas a CAAT box was found in the PPARγ promoter (Fig. 3a). Thus, we proposed PPARγ as potential KLF13 direct target during the process of porcine adipocyte differentiation. Real-time PCR analysis has shown that the PPARγ was coordinately regulated by KLF13 in ASVC (Fig. 2c,e). Furthermore, western blot analysis also confirmed that PPARγ expression was coordinately regulated by KLF13 in ASVC under adipogenic induction condition (Fig. 3b,c).Fig. 3

Bottom Line: Porcine adipocyte differentiation was significantly attenuated by the addition of siRNA against KLF13, whereas overexpression of KLF13 resulted in enhanced porcine adipocyte differentiation.In addition, knockdown and/or overexpression of KLF13 in 3 T3-L1 cells all did not influence expression of PPARγ2.Collectively, our results suggest that KLF13 exist as a key pro-adipogenic transcription factor through transactivating PPARγ expression in porcine adipocyte differentiation, whereas no such effect was detected in mouse adipocyte differentiation.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal Nutrition and Feed Science, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan, P. R. China.

ABSTRACT

Background: Adipogenesis is tightly controlled by a complex network of transcription factors acting at different stages of differentiation. Kruppel-like factors (KLFs) as a family of zinc-finger transcription factors play diverse roles during cell differentiation and development in mammals.

Results: In the present study, we showed that KLF13 acts as a key regulator regulating porcine adipocyte differentiation. The expression of KLF13 was markedly up-regulated during the early stage of porcine adipocyte differentiation, which was followed by expression of PPARγ. Porcine adipocyte differentiation was significantly attenuated by the addition of siRNA against KLF13, whereas overexpression of KLF13 resulted in enhanced porcine adipocyte differentiation. Using promoter deletion and mutation analysis, we identified a KLF13-binding site within -593/-577 region of the porcine PPARγ proximal promoter, indicating that KLF13 directly interacts with porcine PPARγ promoter. However, inhibition of KLF13 by siRNA did not impair mouse adipocyte differentiation. In addition, knockdown and/or overexpression of KLF13 in 3 T3-L1 cells all did not influence expression of PPARγ2.

Conclusions: Collectively, our results suggest that KLF13 exist as a key pro-adipogenic transcription factor through transactivating PPARγ expression in porcine adipocyte differentiation, whereas no such effect was detected in mouse adipocyte differentiation.

No MeSH data available.