2-O-Methylhonokiol Suppresses HCV Replication via TRAF6-Mediated NF-kB Activation
Abstract
:1. Introduction
2. Results
2.1. Inhibitory Effects of 2-O-Methylhonokiol on HCV Replication
2.2. Comparison of 2-O-Methylhonokiol and Honokiol on Cytotoxicity and HCV Replication Inhibition
2.3. 2-O-Methylhonokiol Suppresses HCV Replication through NF-kB Activation
2.4. 2-O-Methylhonokiol Enhances TRAF6-Mediated NF-kB Activation
2.5. 2-O-Methylhonokiol Stimulates Innate Immune Responses
2.6. Conditioned Media from 2-O-Methylhonokiol-Trated Cells also Inhibit HCV Replication
3. Discussion
4. Materials and Methods
4.1. Cells and Compounds
4.2. Plasmids, siRNAs Transfection
4.3. Cell Viability Assay
4.4. Luciferase Reporter Assay
4.5. Quantitative Real-Time Reverse Transcription Polymerase Chain Reaction (qRT-PCR)
4.6. Western Blotting
4.7. Immunoprecipitation
4.8. Nuclear-Cytosol Fraction
4.9. Confocal Immunofluorescence Microscopy
4.10. IFN-α ELISA
4.11. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
HCV | Hepatitis C virus |
NF-kB | Nuclear factor kappa-light-chain-enhancer of activated B cells |
TRAF6 | Tumor necrosis factor receptor (TNFR)-associated factor 6 |
INF | Interferon |
DAA | Direct Acting Antiviral |
PRRs | Pattern Recognition Receptors |
RLRs | Retinoic acid-inducible gene I product (RIG-I)-like receptors |
TLRs | Toll-like receptors |
PAMPs | Pathogen-associated molecular patterns |
LUBAC | Linear ubiquitin chain assembly complex |
PKR | Protein kinase R |
MxA | Myxovirus resistance protein 1 |
CQ | Chloroquine |
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Gene | Direction | Sequence |
---|---|---|
HCV | Forward | GTCTAGCCATGGCGTTAGTATGAG |
Reverse | CTTGTGGTAGCCTGATAGGGT | |
b-Actin | Forward | CATCGAGCACGGCATCGTCA |
Reverse | TCGAAGTCCAGGGCGACATA | |
TRAF6 | Forward | TTTGCTCTTATGGATTGTCCCC |
Reverse | CATTGATGCAGCACAGTTGTC | |
P65 | Forward | CTGTGCGTGTCTCCATGCA |
Reverse | TCGTCTGTATCTGGCAGGTACTG | |
IFN-α | Forward | TTTCTCCTGCCTGAAGGACAG |
Reverse | GCTCATGATTTCTGCTCTGACA | |
IFN-β | Forward | GAACTTTGACATCCCTGAGGAGATTAAGCAGC |
Reverse | GTTCCTTAGGATTTCCACTCTGACTATGGTCC | |
MxA | Forward | AACAACCTGTGCAGCCAGTA |
Reverse | AAGGGCAACTCCTGAGAGTG | |
PKR | Forward | TCTCTGGCGGTCTTCAGAAT |
Reverse | ACTCCCTGCTTCTGACGGTA |
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Jeong, S.; Lee, Y.-s.; Kim, K.; Yoon, J.-s.; Kim, S.; Ha, J.; Kang, I.; Choe, W. 2-O-Methylhonokiol Suppresses HCV Replication via TRAF6-Mediated NF-kB Activation. Int. J. Mol. Sci. 2021, 22, 6499. https://doi.org/10.3390/ijms22126499
Jeong S, Lee Y-s, Kim K, Yoon J-s, Kim S, Ha J, Kang I, Choe W. 2-O-Methylhonokiol Suppresses HCV Replication via TRAF6-Mediated NF-kB Activation. International Journal of Molecular Sciences. 2021; 22(12):6499. https://doi.org/10.3390/ijms22126499
Chicago/Turabian StyleJeong, Suyun, Young-seok Lee, Kiyoon Kim, Ji-su Yoon, Sungsoo Kim, Joohun Ha, Insug Kang, and Wonchae Choe. 2021. "2-O-Methylhonokiol Suppresses HCV Replication via TRAF6-Mediated NF-kB Activation" International Journal of Molecular Sciences 22, no. 12: 6499. https://doi.org/10.3390/ijms22126499