m6A reader IGF2BP1 accelerates apoptosis of high glucose-induced vascular endothelial cells in a m6A-HMGB1 dependent manner

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Biochemistry, Biophysics and Molecular Biology

Main article text

 

Introduction

Materials and Methods

Cell culture and diabetes model treatment

Plasmids construction and cell transfections

RNA extraction and qRT‑PCR

Western blot assay

Cell counting kit‑8 (CCK8) assay

Cellular apoptosis analysis

Ethynyl-2-deoxyuridine (EdU) incorporation assay

m6A quantification assay

RNA immunoprecipitation

m6A methylated RNA immunoprecipitation-PCR

RNA stability

Statistical analysis

Results

IGF2BP1 was up-regulated in HG-induced HUVECs

Knockdown of IGF2BP1 mitigated HG-induced apoptosis of HUVECs

HMGB1 acted as the target of IGF2BP1 by m6A modified sites on HMGB1 mRNA

IGF2BP1 enhanced the stability of HMGB1 mRNA via m6A-dependent manner

Discussion

Supplemental Information

Primer sequences for qRT-PCR and sequences of shRNA.

DOI: 10.7717/peerj.14954/supp-1

Label for WB blot.

DOI: 10.7717/peerj.14954/supp-2

Uncropped blots.

DOI: 10.7717/peerj.14954/supp-3

Additional Information and Declarations

Competing Interests

The authors declare that they have no competing interests.

Author Contributions

Anru Liang conceived and designed the experiments, analyzed the data, authored or reviewed drafts of the article, and approved the final draft.

Jianyu Liu conceived and designed the experiments, prepared figures and/or tables, and approved the final draft.

Yanlin Wei conceived and designed the experiments, prepared figures and/or tables, and approved the final draft.

Yuan Liao conceived and designed the experiments, analyzed the data, authored or reviewed drafts of the article, and approved the final draft.

Fangxiao Wu performed the experiments, prepared figures and/or tables, and approved the final draft.

Jiang Ruan performed the experiments, analyzed the data, prepared figures and/or tables, and approved the final draft.

Junjun Li performed the experiments, analyzed the data, prepared figures and/or tables, and approved the final draft.

Data Availability

The following information was supplied regarding data availability:

The raw data are available as Supplemental Files.

Funding

The authors received no funding for this work.

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