Thymoquinone Suppresses IRF-3-Mediated Expression of Type I Interferons via Suppression of TBK1
Abstract
:1. Introduction
2. Results
2.1. Effect of Thymoquinone on the mRNA Expression of Interferon Genes in LPS-Stimulated RAW264.7 Cells
2.2. Effect of Thymoquinone on the Transcriptional Activation of IRF-3
2.3. Effect of Thymoquinone on Upstream Signaling Events Regulating IRF-3 Activation
3. Discussion
4. Materials and Methods
4.1. Materials
4.2. Cell Culture and Drug Preparation
4.3. Cell Viability Assay
4.4. mRNA Analysis by Semiquantitative Reverse Transcriptase (RT)-Polymerase Chain Reaction (PCR) and Real-Time PCR (qPCR)
4.5. Luciferase Reporter Gene Assay
4.6. Preparation of Whole Cell Lysates and Immunoblotting
4.7. Cellular Thermal Shift Assay (CETSA)
4.8. Statistical Analysis
Author Contributions
Acknowledgments
Conflicts of Interest
References
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PCR Type | Name | Sequence (5′–3′) | |
---|---|---|---|
Reverse Transcriptase PCR | IFN-α | Forward Reverse | CAACACCTACACAGGTTACC AGTGGCTTCCCAGATGTTCC |
IFN-β | Forward Reverse | TCCAAGAAAGGACGAACATT TGAGGACATCTCCCACGTCA | |
GAPDH | Forward Reverse | ACCACAGTCCATGCCATCAC CCACCACCCTGTTGCTGTAG | |
Real Time PCR | IFN-β | Forward Reverse | AAGAGTTACACTGCCTTTGCTATC CACTGTCTGCTGGTGGAGTTCATC |
GAPDH | Forward Reverse | CAATGAATACGGCTACAGCA AGGGAGATGCTCAGTGTTGG |
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Aziz, N.; Son, Y.-J.; Cho, J.Y. Thymoquinone Suppresses IRF-3-Mediated Expression of Type I Interferons via Suppression of TBK1. Int. J. Mol. Sci. 2018, 19, 1355. https://doi.org/10.3390/ijms19051355
Aziz N, Son Y-J, Cho JY. Thymoquinone Suppresses IRF-3-Mediated Expression of Type I Interferons via Suppression of TBK1. International Journal of Molecular Sciences. 2018; 19(5):1355. https://doi.org/10.3390/ijms19051355
Chicago/Turabian StyleAziz, Nur, Young-Jin Son, and Jae Youl Cho. 2018. "Thymoquinone Suppresses IRF-3-Mediated Expression of Type I Interferons via Suppression of TBK1" International Journal of Molecular Sciences 19, no. 5: 1355. https://doi.org/10.3390/ijms19051355