FNDC5 expression closely correlates with muscle fiber types in porcine longissimus dorsi muscle and regulates myosin heavy chains (MyHCs) mRNA expression in C2C12 cells

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Biochemistry, Biophysics and Molecular Biology

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Introduction

Materials & Methods

Animal feeding, muscle sampling, FNDC5 expression, and MyHC mRNA composition analysis

Culture, differentiation, and transfection of C2C12 myoblast cells

ZLN005 or irisin treatment on C2C12 myoblasts

Quantitative real-time PCR (qPCR) and western blotting of cells

Detection of metabolic enzyme activity in cells

Statistical analysis

Results

Knockdown or up expression of FNDC5 gene in C2C12 myoblast cells

Effects of down or up regulating FNDC5 expression on MyHCs mRNA levels in C2C12 myoblast cells

Effects of FNDC5 knockdown expression on PGC1α-FNDC5-MyHCs mRNA expression under ZLN005 in C2C12 myoblast cells

Effects of adding irisin on PGC1α-FNDC5-MyHCs mRNA expression in C2C12 myoblast cells

Effects of adding irisin on metabolic enzyme activities and other downstream gene expression levels in C2C12 myoblast cells

Discussion

Conclusions

Supplemental Information

Raw data of FNDC5mRNA expression and MyHCs mRNA composition in porcine muscle

Each data point indicates the value from one pig.

DOI: 10.7717/peerj.11065/supp-1

Raw data of genes expression or enzymes activity of C2C12 cells in vitro

Each data point indicates the average performance of three repeats measures of cells

DOI: 10.7717/peerj.11065/supp-2

Original picture of WB

DOI: 10.7717/peerj.11065/supp-3

Explanation regarding the double band seen in the Western blot and its the antibody blocking test

DOI: 10.7717/peerj.11065/supp-4

Mycoplasma detection

The instructions of MycAway™ -Color One-Step Mycoplasma Detection Kit and detection results.

DOI: 10.7717/peerj.11065/supp-5

Additional Information and Declarations

Competing Interests

The authors declare there are no competing interests.

Author Contributions

Xiao-Ming Men conceived and designed the experiments, performed the experiments, prepared figures and/or tables, authored or reviewed drafts of the paper, and approved the final draft.

Zi-Wei Xu conceived and designed the experiments, authored or reviewed drafts of the paper, and approved the final draft.

Xin Tao performed the experiments, prepared figures and/or tables, and approved the final draft.

Bo Deng analyzed the data, prepared figures and/or tables, and approved the final draft.

Ke-Ke Qi performed the experiments, authored or reviewed drafts of the paper, and approved the final draft.

Animal Ethics

The following information was supplied relating to ethical approvals (i.e., approving body and any reference numbers):

Laboratory Animal Management Committee of Zhejiang Academy of Agricultural Sciences provided full approval for this research (No. 2018110).

Data Availability

The following information was supplied regarding data availability:

Raw data showing MyHCs mRNA composition in porcine muscle and various genes expressions in C2C12 cells are available in the Supplementary Files.

Funding

This work was supported by grants from the Science and Technology Projects in Zhejiang Province of China (grant numbers 2021C02007, LY17C170005, 2016C02054). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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