METTL16-mediated translation of CIDEA promotes non-alcoholic fatty liver disease progression via m6A-dependent manner

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Biochemistry, Biophysics and Molecular Biology

Main article text

 

Introduction

Materials and Methods

Animal experiments

H&E staining

Oil red O staining

RNA extraction and RNA-seq

qRT-PCR analysis

MeRIP-seq and data analysis

Immunohistochemistry

Cell culture and treatments

METTL16 transfection

Gene silencing with METTL16 siRNA

Western blot

Statistical analysis

Results

Overview of m6A methylation map within mRNA in HFD mice livers

Conjoint analysis of RNA-seq and MeRIP-seq data

GO and KEGG pathway analysis of differentially methylated mRNAs

Both m6A enrichment and expression levels of CIDEA were significantly upregulated in HFD mice livers

The expression levels of METTL16 were substantially increased in HFD mice livers

METTL16-mediated m6A methylation upregulated the expression levels of CIDEA in HepG2 cells

Discussion

Conclusions

Supplemental Information

Primers for reverse transcription quantitative polymerase chain reaction.

DOI: 10.7717/peerj.14379/supp-1

Author Checklist-Full.

DOI: 10.7717/peerj.14379/supp-3

Additional Information and Declarations

Competing Interests

The authors declare that they have no competing interests.

Author Contributions

Jinhong Tang conceived and designed the experiments, performed the experiments, analyzed the data, prepared figures and/or tables, authored or reviewed drafts of the article, and approved the final draft.

Xiangyun Zhao conceived and designed the experiments, performed the experiments, analyzed the data, prepared figures and/or tables, authored or reviewed drafts of the article, and approved the final draft.

Wei Wei performed the experiments, analyzed the data, authored or reviewed drafts of the article, and approved the final draft.

Weiwei Liu performed the experiments, analyzed the data, authored or reviewed drafts of the article, and approved the final draft.

Huining Fan performed the experiments, analyzed the data, authored or reviewed drafts of the article, and approved the final draft.

Xiu ping Liu analyzed the data, authored or reviewed drafts of the article, and approved the final draft.

Yungai Li analyzed the data, authored or reviewed drafts of the article, and approved the final draft.

Long Wang conceived and designed the experiments, performed the experiments, analyzed the data, prepared figures and/or tables, authored or reviewed drafts of the article, and approved the final draft.

Jinghui Guo conceived and designed the experiments, performed the experiments, analyzed the data, prepared figures and/or tables, authored or reviewed drafts of the article, and approved the final draft.

Animal Ethics

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

The animal protocol was approved by the Animal Care and Ethics Committee of Shanghai Jiao Tong University Affiliated Sixth People’s Hospital (IACUC 2020-0035).

DNA Deposition

The following information was supplied regarding the deposition of DNA sequences:

The raw data is available at NCBI GEO: GSE202843, in: https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE202843.

The following secure token has been created to allow review of record GSE202843: kdihieeqrpylzsx.

Data Availability

The following information was supplied regarding data availability:

The following information was supplied regarding data availability: The raw data is available at NCBI GEO, accession numbers: GSE202843, in: https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE202843.

PCR raw data are provided as Supplemental Files.

Funding

The present study was supported by the Science and Technology Innovation Special Fund of Shanghai Jiao Tong University (Grant No. YG2016QN18) and the Scientific Research Fund of Shanghai Jiao Tong University Affiliated Sixth People’s Hospital (Grant No. X-2258). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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