High-Concentrate Feeding to Dairy Cows Induces Apoptosis via the NOD1/Caspase-8 Pathway in Mammary Epithelial Cells
Abstract
1. Introduction
2. Materials and Methods
2.1. Ethics Statement
2.2. Animals, Diets, and Their Trail Plan
2.3. Collection of Samples
2.4. Detection of Apoptosis by the Tunnel Assay
2.5. Isolation of RNA, cDNA Synthesis, and Real-Time (RT-qPCR)
2.6. The Concentration of Caspase-3 and Caspase-8 in Mammary Tissue
2.7. Western Blotting Analysis
2.8. Immunofluorescence Antibody (IFA) Assay
2.9. Statistical Analysis
3. Results
3.1. Determination of Apoptotic Cells in the Mammary Gland Tissue of Dairy Cows
3.2. mRNA Expression of NOD1, Rip-2, Bax, and Bcl2 in Mammary Gland Tissue
3.3. Determination of Caspase-3, Caspase-8, and Caspase-9 mRNA Expressions, and Caspase-3 and Caspase-8 Concentration in Mammary Gland Tissue
3.4. Protein Expression in Mammary Tissue
3.5. Protein Expression and Localization of NOD1 and Caspase-8
4. Discussion
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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Gene | Accession Number | Primer Sequence (5′–3′) | Product Length (bp) | |
---|---|---|---|---|
NOD1 | NM_001256563.1 | Forward | TCAACACTGACCCAGTGAGC | 147 |
Reverse | TGAAGTTGACCAGCTCCACC | |||
β-actin | AY141970 | Forward | CTCTTCCAGCCTTCCTTCCT | 178 |
Reverse | GGGCAGTGATCTCTTTCTGC | |||
Rip-2 | NM_001034610.2 | Forward | ATTCTGGTCCACGGGAGGAGTC | 94 |
Reverse | TCTTGAGGAGCTGACAGGGACC | |||
Caspase-3 | NM_001077840.1 | Forward | CAGCGTCGTAGCTGAACGTAA | 227 |
Reverse | ATCGACAGGCCATGCCAGTAT | |||
Caspase-8 | NM_001045970.2 | Forward | AGCAAATGGTCCAGGCTTTG | 250 |
Reverse | GCTCTTGTTGACCTGCTCAC | |||
Caspase-9 | NM_001205504.1 | Forward | AGCAAATGGTCCAGGCTTTG | 160 |
Reverse | ATTCTCTCGACGGACACAGG | |||
Bcl-2 | NM_001075417-2 | Forward | AGGTTGGTAACCGGACCCTA | 174 |
Reverse | TTCCTGCCTGTCCTCGAATG | |||
Bax | NM_173894.1 | Forward | GCTGTGGACACAGACTCTC | 158 |
Reverse | CTGATCAACTGGGCACCTT |
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ul Aabdin, Z.; Cheng, X.; Dai, H.; Wang, Y.; Sahito, B.; Roy, A.C.; Memon, M.A.; Shen, X. High-Concentrate Feeding to Dairy Cows Induces Apoptosis via the NOD1/Caspase-8 Pathway in Mammary Epithelial Cells. Genes 2020, 11, 107. https://doi.org/10.3390/genes11010107
ul Aabdin Z, Cheng X, Dai H, Wang Y, Sahito B, Roy AC, Memon MA, Shen X. High-Concentrate Feeding to Dairy Cows Induces Apoptosis via the NOD1/Caspase-8 Pathway in Mammary Epithelial Cells. Genes. 2020; 11(1):107. https://doi.org/10.3390/genes11010107
Chicago/Turabian Styleul Aabdin, Zain, Xiaoye Cheng, Hongyu Dai, Yan Wang, Benazir Sahito, Animesh Chandra Roy, Meena Arif Memon, and Xiangzhen Shen. 2020. "High-Concentrate Feeding to Dairy Cows Induces Apoptosis via the NOD1/Caspase-8 Pathway in Mammary Epithelial Cells" Genes 11, no. 1: 107. https://doi.org/10.3390/genes11010107
APA Styleul Aabdin, Z., Cheng, X., Dai, H., Wang, Y., Sahito, B., Roy, A. C., Memon, M. A., & Shen, X. (2020). High-Concentrate Feeding to Dairy Cows Induces Apoptosis via the NOD1/Caspase-8 Pathway in Mammary Epithelial Cells. Genes, 11(1), 107. https://doi.org/10.3390/genes11010107