Humanized L184Q Mutated Surfactant Protein C Gene Alters Alveolar Type 2 Epithelial Cell Fate
Abstract
:1. Introduction
2. Results
2.1. Sftpc L184Q Mutation Altered AT2 Yield and Marker Protein Expression In Vivo
2.2. SftpcL184Q Mutation Downregulated AT2 Lineage In Vitro
2.3. SftpcL184Q Disrupted Organoid Size and Structure
2.4. SftpcL184Q Halted Maturation of Transitional AT2 Cells into AT1 Cells
2.5. SftpcL184Q Did Not Alter CD74, HSP90, or RPS3A1 Expression in Organoids
3. Discussion
4. Materials and Methods
4.1. Animals
4.2. Fluorescence-Activated Cell Sorting (FACS) Mediated AT2 Cell Isolation
4.3. Feeder-Free Culture of Alveolar Organoids
4.4. Western Blotting
4.5. Immunofluorescence Staining
4.6. Flow Cytometry
4.7. EdU Labeling of Proliferating AT2 Cells
4.8. Quantitative Reverse Transcriptase PCR
4.9. Statistics 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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Forward | Reverse | |
---|---|---|
CD74 | GCTGGATGAAGCAGTGGCTCTT | GATGTGGCTGACTTCTTCCTGG |
RPS3A1 | GGTACGATGTGAAAGCTCCAGC | GGCTCACTTCAAACACACGACC |
GAPDH | CATCACTGCCACCCAGAAGACTG | ATGCCAGTGAGCTTCCCGTTCAG |
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Jain, K.G.; Liu, Y.; Zhao, R.; Muire, P.J.; Zhang, J.; Zang, Q.S.; Ji, H.-L. Humanized L184Q Mutated Surfactant Protein C Gene Alters Alveolar Type 2 Epithelial Cell Fate. Int. J. Mol. Sci. 2024, 25, 8723. https://doi.org/10.3390/ijms25168723
Jain KG, Liu Y, Zhao R, Muire PJ, Zhang J, Zang QS, Ji H-L. Humanized L184Q Mutated Surfactant Protein C Gene Alters Alveolar Type 2 Epithelial Cell Fate. International Journal of Molecular Sciences. 2024; 25(16):8723. https://doi.org/10.3390/ijms25168723
Chicago/Turabian StyleJain, Krishan G., Yang Liu, Runzhen Zhao, Preeti J. Muire, Jiwang Zhang, Qun Sophia Zang, and Hong-Long Ji. 2024. "Humanized L184Q Mutated Surfactant Protein C Gene Alters Alveolar Type 2 Epithelial Cell Fate" International Journal of Molecular Sciences 25, no. 16: 8723. https://doi.org/10.3390/ijms25168723