Limits...
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 knockdown or overexpression of KLF13 on the expression of adipogenic factors. a After 2 days transfection of KLF13 siRNA, adipose SV cells were harvested. Real-time PCR was used to determine the mRNA expression of adipogenic factor genes Ebf1, KLF4, C/EBPβ, KLF15, PPARγ and C/EBPα. The protein level was determined by western blotting. Values are represented as mean ± SD. (n = 3). b After 2 days transfection of pcDNA3.1-KLF13, adipose SV cells were harvested. Real-time PCR was used to determine the mRNA expression of adipocyte differentiation-related genes Ebf1, KLF4, C/EBPβ, KLF15, PPARγ and C/EBPα. The protein level was determined by western blotting. Values are represented as mean ± SD. (n = 3). c After 1 days transfection of KLF13 siRNA, adipose SV cells were stimulated in adipogenic induction medium for 2 days. Real-time PCR was used to determine the mRNA expression of adipocyte differentiation-related genes Ebf1, KLF4, C/EBPβ, KLF9, KLF15, PPARγ and C/EBPα. The protein level was determined by western blotting. Values are represented as mean ± SD. (n = 3). d After 1 days transfection of pcDNA3.1-KLF13, adipose SV cells were stimulated in adipogenic induction medium for 2 days. Real-time PCR was used to determine the mRNA expression of KLF13. The protein level was determined by western blotting. Values are represented as mean ± SD. (n = 3). e After 1 days transfection of pcDNA3.1-KLF13, adipose SV cells were stimulated in adipogenic induction medium for 2 days. Real-time PCR was used to determine the mRNA expression of adipocyte differentiation-related genes Ebf1, KLF4, C/EBPβ, KLF15, PPARγ and C/EBPα. Values are represented as mean ± SD. (n = 3) *P < 0.05, **P < 0.01
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4469396&req=5

Fig2: Effect of knockdown or overexpression of KLF13 on the expression of adipogenic factors. a After 2 days transfection of KLF13 siRNA, adipose SV cells were harvested. Real-time PCR was used to determine the mRNA expression of adipogenic factor genes Ebf1, KLF4, C/EBPβ, KLF15, PPARγ and C/EBPα. The protein level was determined by western blotting. Values are represented as mean ± SD. (n = 3). b After 2 days transfection of pcDNA3.1-KLF13, adipose SV cells were harvested. Real-time PCR was used to determine the mRNA expression of adipocyte differentiation-related genes Ebf1, KLF4, C/EBPβ, KLF15, PPARγ and C/EBPα. The protein level was determined by western blotting. Values are represented as mean ± SD. (n = 3). c After 1 days transfection of KLF13 siRNA, adipose SV cells were stimulated in adipogenic induction medium for 2 days. Real-time PCR was used to determine the mRNA expression of adipocyte differentiation-related genes Ebf1, KLF4, C/EBPβ, KLF9, KLF15, PPARγ and C/EBPα. The protein level was determined by western blotting. Values are represented as mean ± SD. (n = 3). d After 1 days transfection of pcDNA3.1-KLF13, adipose SV cells were stimulated in adipogenic induction medium for 2 days. Real-time PCR was used to determine the mRNA expression of KLF13. The protein level was determined by western blotting. Values are represented as mean ± SD. (n = 3). e After 1 days transfection of pcDNA3.1-KLF13, adipose SV cells were stimulated in adipogenic induction medium for 2 days. Real-time PCR was used to determine the mRNA expression of adipocyte differentiation-related genes Ebf1, KLF4, C/EBPβ, KLF15, PPARγ and C/EBPα. Values are represented as mean ± SD. (n = 3) *P < 0.05, **P < 0.01

Mentions: KLF13 is a transcriptional regulator, we sought to identify downstream targets through both gain-of-function and loss-of-function studies under adipogenic or non-adipogenic induction (normal grown medium) conditions. In order to identify bona fide targets of KLF13, we focused on the known key adipogenic regulators that showed coordinate regulation between KLF13 overexpression and knockdown. In the context of normal grown medium, there are four transcription factor genes (Ebf1, KLF15, PPARγ, and C/EBPα) that were significantly diminished by KLF13 knockdown in ASVC (Fig. 2a), while three transcription factor genes (Ebf1, PPARγ, and C/EBPα) whose expression significantly increased upon treatment with KLF13 overexpression in ASVC, C/EBPβ was decreased (Fig. 2b). A total of three transcription factor genes (Ebf1, PPARγ, and C/EBPα) were coordinately regulated by KLF13 in ASVC under normal gown medium.Fig. 2


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 knockdown or overexpression of KLF13 on the expression of adipogenic factors. a After 2 days transfection of KLF13 siRNA, adipose SV cells were harvested. Real-time PCR was used to determine the mRNA expression of adipogenic factor genes Ebf1, KLF4, C/EBPβ, KLF15, PPARγ and C/EBPα. The protein level was determined by western blotting. Values are represented as mean ± SD. (n = 3). b After 2 days transfection of pcDNA3.1-KLF13, adipose SV cells were harvested. Real-time PCR was used to determine the mRNA expression of adipocyte differentiation-related genes Ebf1, KLF4, C/EBPβ, KLF15, PPARγ and C/EBPα. The protein level was determined by western blotting. Values are represented as mean ± SD. (n = 3). c After 1 days transfection of KLF13 siRNA, adipose SV cells were stimulated in adipogenic induction medium for 2 days. Real-time PCR was used to determine the mRNA expression of adipocyte differentiation-related genes Ebf1, KLF4, C/EBPβ, KLF9, KLF15, PPARγ and C/EBPα. The protein level was determined by western blotting. Values are represented as mean ± SD. (n = 3). d After 1 days transfection of pcDNA3.1-KLF13, adipose SV cells were stimulated in adipogenic induction medium for 2 days. Real-time PCR was used to determine the mRNA expression of KLF13. The protein level was determined by western blotting. Values are represented as mean ± SD. (n = 3). e After 1 days transfection of pcDNA3.1-KLF13, adipose SV cells were stimulated in adipogenic induction medium for 2 days. Real-time PCR was used to determine the mRNA expression of adipocyte differentiation-related genes Ebf1, KLF4, C/EBPβ, KLF15, PPARγ and C/EBPα. Values are represented as mean ± SD. (n = 3) *P < 0.05, **P < 0.01
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4469396&req=5

Fig2: Effect of knockdown or overexpression of KLF13 on the expression of adipogenic factors. a After 2 days transfection of KLF13 siRNA, adipose SV cells were harvested. Real-time PCR was used to determine the mRNA expression of adipogenic factor genes Ebf1, KLF4, C/EBPβ, KLF15, PPARγ and C/EBPα. The protein level was determined by western blotting. Values are represented as mean ± SD. (n = 3). b After 2 days transfection of pcDNA3.1-KLF13, adipose SV cells were harvested. Real-time PCR was used to determine the mRNA expression of adipocyte differentiation-related genes Ebf1, KLF4, C/EBPβ, KLF15, PPARγ and C/EBPα. The protein level was determined by western blotting. Values are represented as mean ± SD. (n = 3). c After 1 days transfection of KLF13 siRNA, adipose SV cells were stimulated in adipogenic induction medium for 2 days. Real-time PCR was used to determine the mRNA expression of adipocyte differentiation-related genes Ebf1, KLF4, C/EBPβ, KLF9, KLF15, PPARγ and C/EBPα. The protein level was determined by western blotting. Values are represented as mean ± SD. (n = 3). d After 1 days transfection of pcDNA3.1-KLF13, adipose SV cells were stimulated in adipogenic induction medium for 2 days. Real-time PCR was used to determine the mRNA expression of KLF13. The protein level was determined by western blotting. Values are represented as mean ± SD. (n = 3). e After 1 days transfection of pcDNA3.1-KLF13, adipose SV cells were stimulated in adipogenic induction medium for 2 days. Real-time PCR was used to determine the mRNA expression of adipocyte differentiation-related genes Ebf1, KLF4, C/EBPβ, KLF15, PPARγ and C/EBPα. Values are represented as mean ± SD. (n = 3) *P < 0.05, **P < 0.01
Mentions: KLF13 is a transcriptional regulator, we sought to identify downstream targets through both gain-of-function and loss-of-function studies under adipogenic or non-adipogenic induction (normal grown medium) conditions. In order to identify bona fide targets of KLF13, we focused on the known key adipogenic regulators that showed coordinate regulation between KLF13 overexpression and knockdown. In the context of normal grown medium, there are four transcription factor genes (Ebf1, KLF15, PPARγ, and C/EBPα) that were significantly diminished by KLF13 knockdown in ASVC (Fig. 2a), while three transcription factor genes (Ebf1, PPARγ, and C/EBPα) whose expression significantly increased upon treatment with KLF13 overexpression in ASVC, C/EBPβ was decreased (Fig. 2b). A total of three transcription factor genes (Ebf1, PPARγ, and C/EBPα) were coordinately regulated by KLF13 in ASVC under normal gown medium.Fig. 2

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.