Quantitative Phosphoproteomics Reveals Cell Alignment and Mitochondrial Length Change under Cyclic Stretching in Lung Cells
Abstract
:1. Introduction
2. Results
2.1. Cell Surface Area (CSA) Determination
2.2. Quantitative Phosphoproteome of Lung Cells in Response to Cyclic Stretching
2.3. Functional Enrichment of Stretching-Regulated Phosphoproteins
2.4. Functional Enrichment of Stretching-Regulated Gene Expression
2.5. Uniaxial Cyclic Stretching Resulted in Cell Rearrangement
2.6. Cyclic Stretching Enhanced Mitochondrial Length
2.7. Global View of the Phosphorylation Events Induced by Cyclic Stretching
3. Discussion
4. Materials and Methods
4.1. Cell Cultures
4.2. Selection of Cyclic Stretching Conditions
4.3. Cellular Orientation Measurement
4.4. Protein Extraction
4.5. Phosphoproteome Experiments
4.6. Nano LC-MS/MS Analysis
4.7. Phosphoproteomics Data Processing and Analysis
4.8. Functional Enrichment Analysis of Differential Phosphoproteins
4.9. Transcriptomics Data Analysis
4.10. Immunofluorescence Staining
4.11. Mitochondrial Image Analysis
4.12. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Wang, W.-H.; Hsu, C.-L.; Huang, H.-C.; Juan, H.-F. Quantitative Phosphoproteomics Reveals Cell Alignment and Mitochondrial Length Change under Cyclic Stretching in Lung Cells. Int. J. Mol. Sci. 2020, 21, 4074. https://doi.org/10.3390/ijms21114074
Wang W-H, Hsu C-L, Huang H-C, Juan H-F. Quantitative Phosphoproteomics Reveals Cell Alignment and Mitochondrial Length Change under Cyclic Stretching in Lung Cells. International Journal of Molecular Sciences. 2020; 21(11):4074. https://doi.org/10.3390/ijms21114074
Chicago/Turabian StyleWang, Wei-Hsuan, Chia-Lang Hsu, Hsuan-Cheng Huang, and Hsueh-Fen Juan. 2020. "Quantitative Phosphoproteomics Reveals Cell Alignment and Mitochondrial Length Change under Cyclic Stretching in Lung Cells" International Journal of Molecular Sciences 21, no. 11: 4074. https://doi.org/10.3390/ijms21114074