Integrated Amino Acids and Transcriptome Analysis Reveals Arginine Transporter SLC7A2 Is a Novel Regulator of Myogenic Differentiation
Abstract
:1. Introduction
2. Results
2.1. Amino Acids Levels Exhibit Distinct Changes during C2C12 Myoblast Differentiation
2.2. Amino Acid Metabolism Related Genes Are Enriched during C2C12 Myoblast Differentiation
2.3. Arginine Transporter SLC7A2 Is Upregulated during Myogenic Differentiation
2.4. Knockdown of SLC7A2 Inhibits C2C12 Myoblast Proliferation
2.5. Knockdown of SLC7A2 Inhibits Myoblast Differentiation
2.6. The Expression of SLC7A2 Is Upregulated during Aging
3. Discussion
4. Materials and Methods
4.1. Cell Culture
4.2. RNA Sequencing
4.3. Time-Course Differential Expression Gene Analysis
4.4. RNA Interference
4.5. Quantitative RT-PCR
4.6. Western Blot
4.7. Immunofluorescence Assay
4.8. HPLC Method for Quantification of Intracellular Amino Acid Concentrations in C2C12 Cell
4.9. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Correction Statement
References
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Gene Name | Primer Sequence (Forward) | Primer Sequence (Reverse) |
---|---|---|
Slc7a2 | 5′-GAGTAAGAGGCAGTCACCCG-3′ | 5′-ACTCGCTCTTCAAAGTCGCA-3′ |
Myogenin | 5′-AATGGATTTGGACGCATTGGT-3′ | 5′-TTTGCACTGGTACGTGTTGAT-3′ |
MyoD | 5′-GCACTACAGTGGCGACTCAGAT-3′ | 5′-TAGTAGGCGGTGTCGTAGCCAT-3′ |
Myh1 | 5′-GCGAATCGAGGCTCAGAACAA-3′ | 5′-GTAGTTCCGCCTTCGGTCTTG-3′ |
Myh2 | 5′-CAGAGGCAAGTAGTGGTGGA-3′ | 5′-CAAATTCTCTCTGAACAGGGCA-3′ |
Myh4 | 5′-ACACAGAGTCAGGCGAGTTT-3′ | 5′-CAGTGCGTTCTTGGCCTT-3′ |
Myh7 | 5′-CTTGCTACCCTCAGGTGGCT-3′ | 5′-GAGCCTTGGATTCTCAAACG-3′ |
Gapdh | 5′-TGTCAAGCTCATTTCCTGGT-3′ | 5′-TAGGGCCTCTCTTGCTCAGT-3′ |
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Huang, T.; Zhou, J.; Wang, B.; Wang, X.; Xiao, W.; Yang, M.; Liu, Y.; Wang, Q.; Xiang, Y.; Lan, X. Integrated Amino Acids and Transcriptome Analysis Reveals Arginine Transporter SLC7A2 Is a Novel Regulator of Myogenic Differentiation. Int. J. Mol. Sci. 2024, 25, 95. https://doi.org/10.3390/ijms25010095
Huang T, Zhou J, Wang B, Wang X, Xiao W, Yang M, Liu Y, Wang Q, Xiang Y, Lan X. Integrated Amino Acids and Transcriptome Analysis Reveals Arginine Transporter SLC7A2 Is a Novel Regulator of Myogenic Differentiation. International Journal of Molecular Sciences. 2024; 25(1):95. https://doi.org/10.3390/ijms25010095
Chicago/Turabian StyleHuang, Tiane, Jing Zhou, Benhui Wang, Xiang Wang, Wanli Xiao, Mengqi Yang, Yan Liu, Qiquan Wang, Yang Xiang, and Xinqiang Lan. 2024. "Integrated Amino Acids and Transcriptome Analysis Reveals Arginine Transporter SLC7A2 Is a Novel Regulator of Myogenic Differentiation" International Journal of Molecular Sciences 25, no. 1: 95. https://doi.org/10.3390/ijms25010095