FZD7, Regulated by Non-CpG Methylation, Plays an Important Role in Immature Porcine Sertoli Cell Proliferation
Abstract
:1. Introduction
2. Results
2.1. Characterization of DMRs in Non-CpG Contexts
2.2. Identification of DMGs in Non-CpG Contexts
2.3. The Correlation between DNA Methylation and Gene Expression
2.4. Enrichment Analyses
2.5. Screening of Hub Genes Regulated by Non-CpG Methylation
2.6. Zebularine Treatment
2.7. FZD7 Promotes Sertoli Cell Proliferation
2.8. FZD7 Increases Intracellular Ca2+ Concentrations and Adenosine Triphosphate (ATP) Levels
2.9. FZD7 Induces Early Apoptosis of Sertoli Cells
3. Discussion
4. Materials and Methods
4.1. Ethics Statement
4.2. Data Set from WGBS and RNA-Seq
4.3. Annotation of Gene and Genomic Features
4.4. Identification of DMRs and DMGs in Non-CpG Contexts
4.5. Correlation Analysis of DNA Methylation and Gene Expression
4.6. Enrichment Analysis
4.7. Construction and Module Analysis of the Protein–Protein Interaction (PPI) Network
4.8. Quantitative Real-Time PCR (qPCR)
4.9. Sertoli Cell Culture and Zebularine Treatment
4.10. Bisulfite Sequencing PCR (BSP)
4.11. RNA Interference (RNAi) of FZD7 and Transfection
4.12. Immunofluorescence Staining
4.13. Cell Cycle Assay
4.14. Cell Proliferation Assay
4.15. Cell Apoptosis Assay
4.16. Western Blotting
4.17. Measurement of the Intracellular Ca2+ Concentration
4.18. Adenosine Triphosphate (ATP) Assay
4.19. Prostaglandin E2 (PGE2) Assay
4.20. Apoptosis-Medium Treatment
4.21. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Rank | Gene Symbol | Score |
---|---|---|
1 | VEGFA | 36 |
2 | PECAM1 | 21 |
3 | FZD7 | 17 |
4 | CXCL12 | 16 |
5 | VANGL1 | 15 |
6 | WNT9A | 14 |
6 | GRAP | 14 |
8 | FGF18 | 12 |
8 | ITGB6 | 12 |
10 | RYK | 10 |
Genes | Zebularine-Treated Group | DMSO-Treated Group | ||||
---|---|---|---|---|---|---|
CpG | CHG | CHH | CpG | CHG | CHH | |
FZD7 | 95.62 | 0 | 0.71 | 96.88 | 1.36 | 1.07 |
IFITM1 | 25 | 0 | 0 | 30 | 0 | 0.18 |
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Yang, A.; Yan, S.; Yin, Y.; Chen, C.; Tang, X.; Ran, M.; Chen, B. FZD7, Regulated by Non-CpG Methylation, Plays an Important Role in Immature Porcine Sertoli Cell Proliferation. Int. J. Mol. Sci. 2023, 24, 6179. https://doi.org/10.3390/ijms24076179
Yang A, Yan S, Yin Y, Chen C, Tang X, Ran M, Chen B. FZD7, Regulated by Non-CpG Methylation, Plays an Important Role in Immature Porcine Sertoli Cell Proliferation. International Journal of Molecular Sciences. 2023; 24(7):6179. https://doi.org/10.3390/ijms24076179
Chicago/Turabian StyleYang, Anqi, Saina Yan, Yanfei Yin, Chujie Chen, Xiangwei Tang, Maoliang Ran, and Bin Chen. 2023. "FZD7, Regulated by Non-CpG Methylation, Plays an Important Role in Immature Porcine Sertoli Cell Proliferation" International Journal of Molecular Sciences 24, no. 7: 6179. https://doi.org/10.3390/ijms24076179