The Complement Component 4 Binding Protein α Gene: A Versatile Immune Gene That Influences Lipid Metabolism in Bovine Mammary Epithelial Cell Lines
Abstract
:1. Introduction
2. Results
2.1. Construction of C4BPA Knockout BMECs Line Model
2.2. The Proliferation Rate and Apoptosis Rate of KO-BMECs and WT Cell Line
2.3. Lipid Accumulation in BMECs of C4BPA Gene KO or WT Cell Line
2.4. Triacylglycerol (TG), Cholesterol (CHOL), and Free Fatty Acid (FFA) Contents in BMECs of C4BPA Gene KO or WT Cell Line
2.5. Results of Fatty Acid Contents in BMECs of C4BPA Gene KO or WT Cell Line Based on the GC-MS
2.6. Identification of DEGs in BMECs of C4BPA Gene KO or WT Cell Line
2.7. Functional Enrichment Analysis of the DEGs in BMECs of C4BPA Gene KO or WT Cell Line
2.8. Expression of Different Genes Related to Fat Metabolism
3. Discussion
4. Materials and Methods
4.1. Cell Culture and the Construction of C4BPA Knockout BMECs Line Model
4.2. Construction of C4BPA Gene BMECs KO Cell Line
4.3. Analysis of mRNA and Protein Levels in BMECs of C4BPA Gene KO or WT Cell Line
4.4. Cell Apoptosis in BMECs of C4BPA Gene KO or WT Cell Line
4.5. Lipid Droplet Accumulation in BMECs of C4BPA Gene KO or WT Cell Line
4.6. Triglycerides, Cholesterol, and FFA Content in BMECs of C4BPA Gene KO or WT Cell Line
4.7. Intracellular Fatty Acids Content in BMECs of C4BPA Gene KO or WT Cell Line
4.8. GC-MS Analysis in BMECs of C4BPA Gene KO or WT Cell Line
4.9. DEGs and RNA Sequencing Mapping in BMECs of C4BPA Gene KO or WT Cell Line
4.10. GO and KEGG Analysis of DEGs
4.11. Western Blotting
4.12. Statistical Analysis
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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SgRNA Strand | Target-Seq | Location | Gene ID |
---|---|---|---|
C4BPA-sgRNA1-F | CACCGAGAGAGCTGATGGATCATTG | Intron 3 | 281651 |
C4BPA-sgRNA1-R | AAACCAATGATCCATCAGCTCTCTC | Intron 3 | 281651 |
C4BPA-sgRNA2-F | CACCGAGTGTATCCCTGTTTAAAGA | Intron 5 | 281651 |
C4BPA-sgRNA2-R | AAACTCTTTAAACAGGGATACACTC | Intron 5 | 281651 |
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Chen, X.; Zhao, Z.; Jiang, X.; Li, J.; Miao, F.; Yu, H.; Lin, Z.; Jiang, P. The Complement Component 4 Binding Protein α Gene: A Versatile Immune Gene That Influences Lipid Metabolism in Bovine Mammary Epithelial Cell Lines. Int. J. Mol. Sci. 2024, 25, 2375. https://doi.org/10.3390/ijms25042375
Chen X, Zhao Z, Jiang X, Li J, Miao F, Yu H, Lin Z, Jiang P. The Complement Component 4 Binding Protein α Gene: A Versatile Immune Gene That Influences Lipid Metabolism in Bovine Mammary Epithelial Cell Lines. International Journal of Molecular Sciences. 2024; 25(4):2375. https://doi.org/10.3390/ijms25042375
Chicago/Turabian StyleChen, Xuanxu, Zhihui Zhao, Xinyi Jiang, Jing Li, Fengshuai Miao, Haibin Yu, Ziwei Lin, and Ping Jiang. 2024. "The Complement Component 4 Binding Protein α Gene: A Versatile Immune Gene That Influences Lipid Metabolism in Bovine Mammary Epithelial Cell Lines" International Journal of Molecular Sciences 25, no. 4: 2375. https://doi.org/10.3390/ijms25042375