The High Level of RANKL Improves IκB/p65/Cyclin D1 Expression and Decreases p-Stat5 Expression in Firm Udder of Dairy Goats
Abstract
:1. Introduction
2. Results
2.1. The Firm Udder Structure of Guanzhong Dairy Goats
2.2. Hormones Serum Levels and Receptor Mammary Expression in Goats with Firm Udder
2.3. Transcriptome Sequencing and Validation of Mammary Gland in Guanzhong Dairy Goats with Firm Udder
2.4. The Effects of High RANKL Level on Primary GMECs Proliferation, Function and Formatting Acini
2.5. The Effects of GMECs and Adipocyte-like Cells Co-Culturing on IκB/p65/Cyclin D1 and Elf5 in GMECs
3. Discussion
4. Materials and Methods
4.1. Chemicals and Guanzhong Dairy Goat
4.2. Paraffin Embedment, Hematoxylin-Eosin (HE) and IHC Staining of Goat Mammary Gland
4.3. Transmission Electron Microscope Sample Preparation and Dyeing
4.4. RNA Extraction SOP for Mammary Gland
4.5. RNA-Seq Library Preparation Protocol (DNBSEQ)
4.6. Determination of Serum E2, PROG, PRL, RANKL and OPG Levels in Dairy Goats with Firm Udder
4.7. Primary Culture and RANKL/OPG Treatment of Goat Mammary Epithelial Cells (GMECs)
4.8. Three-Dimensional Culture of GMECs
4.9. Isolation and Culture of Adipose Derived Stem Cells (ADSCs) from Goat Mammary Gland
4.10. Induction of Adipogenic Differentiation of ADSCs from Goat Mammary Gland
4.11. Induction of Osteogenesis Differentiation of ADSCs from Goat Mammary Gland
4.12. Induction of Chondrogenesis Differentiation of ADSCs from Goat Mammary Gland
4.13. Co-Culture of GMECs and Adipocyte-like Cells Induced by ADSCs
4.14. Total Protein Extraction and Western Blotting
4.15. RNA Isolation and Quantitative Real-Time PCR (qRT-PCR) Analysis
4.16. Immunofluorescence Staining
4.17. Statistical Analysis
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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Gao, Z.; Shao, D.; Zhao, C.; Liu, H.; Zhao, X.; Wei, Q.; Ma, B. The High Level of RANKL Improves IκB/p65/Cyclin D1 Expression and Decreases p-Stat5 Expression in Firm Udder of Dairy Goats. Int. J. Mol. Sci. 2023, 24, 8841. https://doi.org/10.3390/ijms24108841
Gao Z, Shao D, Zhao C, Liu H, Zhao X, Wei Q, Ma B. The High Level of RANKL Improves IκB/p65/Cyclin D1 Expression and Decreases p-Stat5 Expression in Firm Udder of Dairy Goats. International Journal of Molecular Sciences. 2023; 24(10):8841. https://doi.org/10.3390/ijms24108841
Chicago/Turabian StyleGao, Zhen, Dan Shao, Chunrui Zhao, Haokun Liu, Xiaoe Zhao, Qiang Wei, and Baohua Ma. 2023. "The High Level of RANKL Improves IκB/p65/Cyclin D1 Expression and Decreases p-Stat5 Expression in Firm Udder of Dairy Goats" International Journal of Molecular Sciences 24, no. 10: 8841. https://doi.org/10.3390/ijms24108841