Expression of Interferons Lambda 3 and 4 Induces Identical Response in Human Liver Cell Lines Depending Exclusively on Canonical Signaling
Abstract
:1. Introduction
2. Results
2.1. Transient Transfection with Tagged and Non-tagged IFNL3 and IFNL4 Induced Similar mRNA Levels of Interferons but Different Levels of Interferon-Stimulated Genes
2.2. Cytosolic Expression of IFNL3 or IFNL4 Did Not Modify JAK-STAT Signaling
2.3. IFNL3 and IFNL4 Induced Identical Transcriptome Response in Wild Type Cells
2.4. IFNLR1 and IL10R2 Deficiency Abolished JAK-STAT Signaling by Both IFNL3 and IFNL4
3. Discussion
4. Materials and Methods
4.1. Chemicals and Antibodies
4.2. Cell Culture
4.3. Generation of Knockout Cell Lines
4.4. Expression Constructs
4.5. Transient Transfection
4.6. Interferon Treatment
4.7. Real time PCR Analysis
4.8. RNA Sequencing and Transcriptome Analysis
4.9. Immunoblotting
4.10. Immunofluorescence Studies
4.11. Quantification of IFNLs
4.12. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
ISGs | Interferon stimulated genes |
IFNL3 | Interferons lambda 3 |
IFNL4 | Interferons lambda 4 |
IFNLR1 | Interferon lambda receptor 1 |
IL10R2 | Interleukin 10 receptor 2 |
JAK1 | Tyrosine-protein kinase JAK1 |
JAK2 | Tyrosine-protein kinase JAK2 |
TYK2 | Non-receptor tyrosine-protein kinase TYK2 |
STAT | Signal transducer and activator of transcription |
IRF-9 | Interferon regulatory factor 9 |
ISGF-3 | Interferon Stimulated Gene Factor 3 |
HCV | Hepatitis C virus |
MYC | Myc proto-oncogene protein |
CRISPR | Clustered regularly interspaced short palindromic repeats |
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Lunova, M.; Kubovciak, J.; Smolková, B.; Uzhytchak, M.; Michalova, K.; Dejneka, A.; Strnad, P.; Lunov, O.; Jirsa, M. Expression of Interferons Lambda 3 and 4 Induces Identical Response in Human Liver Cell Lines Depending Exclusively on Canonical Signaling. Int. J. Mol. Sci. 2021, 22, 2560. https://doi.org/10.3390/ijms22052560
Lunova M, Kubovciak J, Smolková B, Uzhytchak M, Michalova K, Dejneka A, Strnad P, Lunov O, Jirsa M. Expression of Interferons Lambda 3 and 4 Induces Identical Response in Human Liver Cell Lines Depending Exclusively on Canonical Signaling. International Journal of Molecular Sciences. 2021; 22(5):2560. https://doi.org/10.3390/ijms22052560
Chicago/Turabian StyleLunova, Mariia, Jan Kubovciak, Barbora Smolková, Mariia Uzhytchak, Kyra Michalova, Alexandr Dejneka, Pavel Strnad, Oleg Lunov, and Milan Jirsa. 2021. "Expression of Interferons Lambda 3 and 4 Induces Identical Response in Human Liver Cell Lines Depending Exclusively on Canonical Signaling" International Journal of Molecular Sciences 22, no. 5: 2560. https://doi.org/10.3390/ijms22052560