Knockdown of both FoxO1 and C/EBPβ promotes adipogenesis in porcine preadipocytes through feedback regulation

Authors

  • Xiong Yan,

    1. Laboratory of Animal Fat Deposition and Muscle Development, College of Animal Science and Technology, Northwest A&F University, Yangling, China
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  • Pang Weijun,

    Corresponding author
    • Laboratory of Animal Fat Deposition and Muscle Development, College of Animal Science and Technology, Northwest A&F University, Yangling, China
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  • Wei Ning,

    1. Laboratory of Animal Fat Deposition and Muscle Development, College of Animal Science and Technology, Northwest A&F University, Yangling, China
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  • Wang Yu,

    1. Laboratory of Animal Fat Deposition and Muscle Development, College of Animal Science and Technology, Northwest A&F University, Yangling, China
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  • Ren Wenkai,

    1. Scientific Observing and Experimental Station of Animal Nutrition and Feed Science in South-Central, Ministry of Agriculture, Hunan Provincial Engineering Research Center of Healthy Livestock, Key Laboratory of Agro-Ecological Processes in Subtropical Region, Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha, Hunan, China
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  • Yang Gongshe

    1. Laboratory of Animal Fat Deposition and Muscle Development, College of Animal Science and Technology, Northwest A&F University, Yangling, China
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Corresponding author: e-mail: pwj1226@nwsuaf.edu.cn

Abstract

FoxO1 and C/EBPβ are important transcription factors during adipogenic differentiation; however, the potential mechanisms of their inter-regulation in adipogenesis remain unknown. We found that FoxO1 and C/EBPβ are abundant in adipose tissues of 3- and 180-day pigs at the mRNA and protein levels. During porcine preadipocyte differentiation, C/EBPβ expression increases to the peak at Day 2 and then decreases. In contrast, FoxO1 is lowest on Day 2 and gradually increases. Knockdown of FoxO1 or C/EBPβ with lentivirus-mediated shRNA enhances or inhibits lipid accumulation and adipogenic maker (C/EBPα and aP2) expression, respectively. FoxO1 depletion causes a mild decrease of C/EBPβ protein level, and C/EBPβ interference also inhibits the expression of FoxO1 protein. Knockdown of both FoxO1 and C/EBPβ promotes lipid accumulation at Day 8, and increases the adipogenic markers during differentiation in comparison with the controls and the FoxO1 knockdown alone. Co-immunoprecipitation experiments indicate that FoxO1 binds to C/EBPβ in adipose tissues and in vitro. In conclusion, the results suggest that FoxO1 and C/EBPβ regulate preadipocyte adipogenesis possibly through C/EBPβ → FoxO1 → C/EBPβ feedback regulatory loop and FoxO1-C/EBPβ protein complex.

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