Porcine Epidemic Diarrhea Virus Induces Autophagy to Benefit Its Replication
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cells and Viruses
2.2. Virus Infection
2.3. Quantitative Real-Time PCR
2.4. Transmission Electron Microscopy
2.5. Confocal Fluorescence Microscopy
2.6. Western Blot Analysis
2.7. RNA Interference
2.8. Cell Viability Assay
2.9. Statistical Analysis
3. Results
3.1. PEDV Infection Increases the Levels of Autophagy in Vero Cells
3.2. PEDV Infection Can Enhance Autophagy Flux
3.3. Pharmacological Inhibition of Autophagy Decreases Viral Yield
3.4. Silencing Endogenous Beclin1 or ATG5 Gene Reduces the PEDV Titer
3.5. Autophagy Has a Positive Correlation with NF-κB Signaling Pathway
3.6. Pharmacological Regulation of Autophagy Does Not Affect Cell Viability
4. Discussion
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Primer | Sequence (5’–3’) |
---|---|
PEDV-F | CGTACAGGTAAGTCAATTAC |
PEDV-R | GATGAAGCATTGACTGAA |
PEDV-probe-M | TTCGTCACAGTCGCCAAGG |
ATG5-F | TTCACGCTATATCAGGAT |
ATG5-R | ATCTCACTAATGTCTTCTTG |
Beclin1-F | TGGCACAATCAATAACTTC |
Beclin1-R | CAAGCAGCATTAATCTCAT |
IL-6-F | TGTGAAAGCAGCAAAGAG |
IL-6-R | AGTGTCCTCATTGAATCCA |
IL-1β-F | GCGGCAACGAGGATGACTT |
IL-1β-R | TGGCTACAACAACTGACACGG |
IL-8-F | GGAACCATCTCGCTCTGTGTAA |
IL-8-R | GGTCCACTCTCAATCACTCTCAG |
CCL5-F | ACGCCTCGCTGTCATCCT |
CCL5-R | GCACTTGCCACTGGTGTAGAA |
TNF-α-F | CACCACGCTCTTCTGTCT |
TNF-α-R | AGATGATCTGACTGCCTGAG |
MCP-1-F | CTTCTGTGCCTGCTGCTCATA |
MCP-1-R | ACTTGCTGCTGGTGATTCTTCT |
GAPDH-F | ACATCATCCCTGCCTCTACTG |
GAPDH-R | CCTGCTTCACCACCTTCTTG |
β-actin-F | TTAGTTGCGTTACACCCTTTC |
β-actin-R | ACCTTCACCGTTCCAGTT |
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Guo, X.; Zhang, M.; Zhang, X.; Tan, X.; Guo, H.; Zeng, W.; Yan, G.; Memon, A.M.; Li, Z.; Zhu, Y.; et al. Porcine Epidemic Diarrhea Virus Induces Autophagy to Benefit Its Replication. Viruses 2017, 9, 53. https://doi.org/10.3390/v9030053
Guo X, Zhang M, Zhang X, Tan X, Guo H, Zeng W, Yan G, Memon AM, Li Z, Zhu Y, et al. Porcine Epidemic Diarrhea Virus Induces Autophagy to Benefit Its Replication. Viruses. 2017; 9(3):53. https://doi.org/10.3390/v9030053
Chicago/Turabian StyleGuo, Xiaozhen, Mengjia Zhang, Xiaoqian Zhang, Xin Tan, Hengke Guo, Wei Zeng, Guokai Yan, Atta Muhammad Memon, Zhonghua Li, Yinxing Zhu, and et al. 2017. "Porcine Epidemic Diarrhea Virus Induces Autophagy to Benefit Its Replication" Viruses 9, no. 3: 53. https://doi.org/10.3390/v9030053