Lysine Deprivation Suppresses Adipogenesis in 3T3-L1 Cells: A Transcriptome Analysis
Abstract
:1. Introduction
2. Results
2.1. Lysine Deprivation Suppressed Adipogenesis
2.2. Transcriptome Profile of the 3T3-L1 Cells
2.3. Comparison between Differentiated and Undifferentiated 3T3-L1 Cells
2.4. Identification of Lysine-Depletion-Induced Pathways
2.5. Genes Critical for Inhibition of 3T3-L1 Cell Differentiation and Adipogenesis
2.6. Pathways That Could Be Altered by Differentiation Medium but Not Critical for the Adipogenesis Process
2.7. Suppression of Adipogenesis Is Not Limited to Lysine Depletion but Also Applied to Methionine and Cystine Deprivation Partially via IL6 Overexpression
3. Discussion
4. Materials and Methods
4.1. Cell Culture
4.2. RNA Extraction, cDNA Synthesis, and RNA Sequencing
4.3. Differentially Expressed GENES (DEG) Analysis
4.4. Functional Annotation Analysis
4.5. Integration of Protein–Protein Interaction (PPI) Network and Module Analysis
4.6. Real-Time PCR Analysis
4.7. Oil-Red O (ORO) Staining
4.8. Medium Amino Acid Analysis
4.9. Western Blotting
4.10. Agilent Seahorse XF ATP Real-Time Rate Assay
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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Gene | Forward | Reverse |
---|---|---|
FASn | CCCAGCGGTAGAGAATAGCA | GGGTCCACTAAACTGAGCCT |
SCD1 | TCCAACTCATGTGCCTCTGT | AACAACCAACCCTCGCATTC |
SREBF1 | TTGTTCCTTTGCCTTCCAGC | GATGCCGACCAGATTCCCTA |
PPARg | TGACAGACCTCAGGCAGATC | AGAAGGAACACGTTGTCAGC |
IL6 | TCCTTCCTACCCCAATTTCCA | GTCCACAAACTGATATGCTTAGG |
Tbp | TGCTGTTGGTGATTGTTGGT | ACTGGGAAGGCGGAATGTAT |
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Lee, L.M.-Y.; Lin, Z.-Q.; Zheng, L.-X.; Tu, Y.-F.; So, Y.-H.; Zheng, X.-H.; Feng, T.-J.; Wang, X.-Y.; Wong, W.-T.; Leung, Y.-C. Lysine Deprivation Suppresses Adipogenesis in 3T3-L1 Cells: A Transcriptome Analysis. Int. J. Mol. Sci. 2023, 24, 9402. https://doi.org/10.3390/ijms24119402
Lee LM-Y, Lin Z-Q, Zheng L-X, Tu Y-F, So Y-H, Zheng X-H, Feng T-J, Wang X-Y, Wong W-T, Leung Y-C. Lysine Deprivation Suppresses Adipogenesis in 3T3-L1 Cells: A Transcriptome Analysis. International Journal of Molecular Sciences. 2023; 24(11):9402. https://doi.org/10.3390/ijms24119402
Chicago/Turabian StyleLee, Leo Man-Yuen, Zhi-Qiang Lin, Lu-Xi Zheng, Yi-Fan Tu, Yik-Hing So, Xiu-Hua Zheng, Tie-Jun Feng, Xi-Yue Wang, Wai-Ting Wong, and Yun-Chung Leung. 2023. "Lysine Deprivation Suppresses Adipogenesis in 3T3-L1 Cells: A Transcriptome Analysis" International Journal of Molecular Sciences 24, no. 11: 9402. https://doi.org/10.3390/ijms24119402