miR-143 Targeting CUX1 to Regulate Proliferation of Dermal Papilla Cells in Hu Sheep
Abstract
:1. Introduction
2. Materials and Methods
2.1. Skin Tissue Collection, RNA and Protein Extraction
2.2. Cell Culture and Transfection
2.3. Cell Proliferation and Cell Cycle
2.4. Target Gene Prediction for miR-143
2.5. Quantitative RT-PCR and Western Blot
2.6. Luciferase Reporter Assay
2.7. Statistical Analysis
3. Results
3.1. miR-143 Promoting Dermal Papilla Cells Proliferation
3.2. Target Genes Prediction of miR-143
3.3. CUX1 and KRT71 Highly Expressed in Small Waves Group
3.4. CUX1 and KRT71 Targeting to miR-143
4. Discussion
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Gene/miRNA Name | Forward Primer (5′-3′) | Reverse Primer (5′-3′) | Product Length (bp) |
---|---|---|---|
CUX1 | GCACGACATTGAGACGGAG | AGCTATGGTCTCAGCCTGGT | 160 |
KRT71 | GGCTCATCCAGAGAATCCGC | GAGCATTGTCACCCCTCTGT | 102 |
GAPDH | TCTCAAGGGCATTCTAGGCTAC | GCCGAATTCATTGTCGTACCAG | 151 |
CDK2 | AGAAGTGGCTGCATCACAAG | TCTCAGAATCTCCAGGGAATAG | 92 |
PCNA | CGAGGGCTTCGACACTTAC | GTCTTCATTGCCAGCACATT | 97 |
Cyclin d1 | CCGAGGAGAACAAGCAGATC | GAGGGTGGGTTGGAAATG | 91 |
miR-143 | CGCGTGAGATGAAGCACTG | AGTGCAGGGTCCGAGGTATT | variable |
U6 | CTCGCTTCGGCAGCACA | AACGCTTCACGAATTTGCGT | 95 |
Gene Name | Primer | Forward Primer (5′-3′) |
---|---|---|
CUX1 | CUX1-WT-F | CCctcgagCCATCTCCCTTTAAAGAGGG |
CUX1-WI-R | TTgcggccgcGACTTAGAGCTCGCTGTCC | |
KRT71 | KRT71-WT-F | CCctcgagTTGTCCCAGCTCCTGCTT |
KRT71-WT-R | TTgcggccgcTATTGAGGATGCAG | |
CUX1 | CUX1-MUT1-F | CTTGGGGACAGAGACCTGTGAGCTGTGACACTG |
CUX1-MUT1-R | CAGTGTCACAGCTCACAGGTCTCTGTCCCCAAG | |
KRT71 | KRT71-MUT1-F | GGGAAGAGGAGGCCCCATCTTTTCTCCTTTC |
KRT71-MUT1-R | GAAAGGAGAAAAGATGGGGCCTCCTCTTCCC |
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Hu, T.; Huang, S.; Lv, X.; Wang, S.; Getachew, T.; Mwacharo, J.M.; Haile, A.; Sun, W. miR-143 Targeting CUX1 to Regulate Proliferation of Dermal Papilla Cells in Hu Sheep. Genes 2021, 12, 2017. https://0-doi-org.brum.beds.ac.uk/10.3390/genes12122017
Hu T, Huang S, Lv X, Wang S, Getachew T, Mwacharo JM, Haile A, Sun W. miR-143 Targeting CUX1 to Regulate Proliferation of Dermal Papilla Cells in Hu Sheep. Genes. 2021; 12(12):2017. https://0-doi-org.brum.beds.ac.uk/10.3390/genes12122017
Chicago/Turabian StyleHu, Tingyan, Sainan Huang, Xiaoyang Lv, Shanhe Wang, Tesfaye Getachew, Joram M. Mwacharo, Aynalem Haile, and Wei Sun. 2021. "miR-143 Targeting CUX1 to Regulate Proliferation of Dermal Papilla Cells in Hu Sheep" Genes 12, no. 12: 2017. https://0-doi-org.brum.beds.ac.uk/10.3390/genes12122017