CircRNA-5335 Regulates the Differentiation and Proliferation of Sheep Preadipocyte via the miR-125a-3p/STAT3 Pathway
Abstract
:Simple Summary
Abstract
1. Introduction
1.1. Intramuscular Fat Accumulation and Preadipocytes in Sheep
1.2. IMF Regulatory Factors and Pathways
1.3. CircRNA and IMF
2. Materials and Methods
2.1. Cell Separation and Culture
2.2. Dual-Luciferase Activity Assay
2.3. Identification of circRNA
2.4. Preadipocyte Differentiation
2.5. Oil Red O(ORO) Staining
2.6. Cell Proliferation Assay
2.7. Real-Time Quantitative PCR (RT-qPCR)
2.8. Western Blotting (WB)
2.9. Statistical Analysis
3. Results
3.1. Expression Levels of circRNA-5335/miR-125a-3p/STAT3 during Sheep Preadipocyte Differentiation
3.2. Knockdown of STAT3 Inhibits Differentiation Process of Preadipocytes
3.3. STAT3 Prevents Preadipocytes from Multiplying
3.4. miR-125a-3p Regulates the Differentiation of Preadipocytes through STAT3 Signaling Pathway
3.5. miR-125a-3p Inhibits the Proliferation of Preadipocytes in Sheep
3.6. CircRNA-5335 Regulates the Expression of STAT3 by Binding to miR-125a-3p
3.7. circRNA-5335/miR-125a-3p/STAT3 Regulates the Differentiation and Proliferation of Preadipocytes
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Target Gene | Primer Sequence (5′-3′) | Product Size (bp) | Accession Number |
---|---|---|---|
STAT3 | F:5′-GGAACCTTACACCAAACAGCA-3′ R:5′-AGGGTAGAGATAGACCAGCGG-3′ | 115 | XM_042255967.1 |
PCNA | F:5′-GTGGAGAACTTGGAAATGGAA-3′ R:5′-GAGACAGTGGAGTGGCTTATG-3′ | 153 | XM_004014340.5 |
PPARG | F:5′-CATTTCTGCTCCGCACTAC-3′ R:5′-TGGAACCCTGACGCTTT-3′ | 234 | NM_001100921.1 |
CEBPA | F:5′-GTGGAGACGCAACAGAAGGT-3′ R:5′-AGTTCGCGGCTCAGTTGTT-3′ | 83 | NM_001308574.1 |
GAPDH | F:5′-AAGTTCAACGGCACAGTCA-3′ R:5′-ACCACATACTCAGCACCAGC-3′ | 125 | NM_001190390.1 |
Target Gene | Primer Sequence (5′-3′) |
---|---|
CircRNA-5335 convergent | F: CGACAAAGAGGAAATAGCAAT R: ACAGGTGATGTTCTAAAGGAG |
CircRNA-5335 divergent | F: CACACTCTTGGATTCAGCAGC R: CCTCTTTGATAGGACACTCGT |
miRNA-125a-3p | F: CGCGACAGGTGAGGTTCTT R: AGTGCAGGGTCCGAGGTATT RT:GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACGCTCCC |
U6 | F: GGAACGATACAGAGAAGATTAGC R: TGGAACGCTTCACGAATTTGCG |
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Guo, W.; Ciwang, R.; Wang, L.; Zhang, S.; Liu, N.; Zhao, J.; Zhou, L.; Li, H.; Gao, X.; He, J. CircRNA-5335 Regulates the Differentiation and Proliferation of Sheep Preadipocyte via the miR-125a-3p/STAT3 Pathway. Vet. Sci. 2024, 11, 70. https://doi.org/10.3390/vetsci11020070
Guo W, Ciwang R, Wang L, Zhang S, Liu N, Zhao J, Zhou L, Li H, Gao X, He J. CircRNA-5335 Regulates the Differentiation and Proliferation of Sheep Preadipocyte via the miR-125a-3p/STAT3 Pathway. Veterinary Sciences. 2024; 11(2):70. https://doi.org/10.3390/vetsci11020070
Chicago/Turabian StyleGuo, Wei, Renzeng Ciwang, Lei Wang, Shuer Zhang, Nan Liu, Jinshan Zhao, Lisheng Zhou, Hegang Li, Xiaoxiao Gao, and Jianning He. 2024. "CircRNA-5335 Regulates the Differentiation and Proliferation of Sheep Preadipocyte via the miR-125a-3p/STAT3 Pathway" Veterinary Sciences 11, no. 2: 70. https://doi.org/10.3390/vetsci11020070