An Integrative Transcriptome Subtraction Strategy to Identify Human lncRNAs That Specifically Play a Role in Activation of Human Hepatic Stellate Cells
Abstract
:1. Introduction
2. Results
2.1. Identifying an In Vitro Culture System That Is Suitable to Study hHSC Activation
2.2. Identification of VSMC Transdifferentiation as a Functionally Relevant System That Shares the Key Elements of Fibrogenesis with hHSC Activation
2.3. Bioinformatic Screening of lncRNAs Functioning in HSC/VSMC Transdifferentiation
2.4. CARMN Expression in Stellate Cells Is Induced by TGFβ
2.5. Loss of Function of CARMN Attenuates HSC Activation
2.6. Gain of Function of CARMN Augments HSC Activation
3. Discussion
4. Materials and Methods
4.1. Culture and Treatment of Human HSC
4.2. Immunofluorescence Staining
4.3. RNA Isolation and RT-qPCR
4.4. RNA Sequencing and Bioinformatics Analysis
4.5. Treatment of LX-2 Cells with CARMN Knockdown
4.6. Rapid Amplification of cDNA Ends (RACE)
4.7. Cellular Fractionation
4.8. Western Blot
4.9. Collagen Staining
4.10. Wound Healing Assay
4.11. EdU Assay
4.12. Adenovirus Production and LX-2 Infection
4.13. CRISPR Activation
4.14. Statistics
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Ma, Y.; Harris, J.; Li, P.; Jiang, C.; Sun, H.; Cao, H. An Integrative Transcriptome Subtraction Strategy to Identify Human lncRNAs That Specifically Play a Role in Activation of Human Hepatic Stellate Cells. Non-Coding RNA 2024, 10, 34. https://doi.org/10.3390/ncrna10030034
Ma Y, Harris J, Li P, Jiang C, Sun H, Cao H. An Integrative Transcriptome Subtraction Strategy to Identify Human lncRNAs That Specifically Play a Role in Activation of Human Hepatic Stellate Cells. Non-Coding RNA. 2024; 10(3):34. https://doi.org/10.3390/ncrna10030034
Chicago/Turabian StyleMa, Yonghe, Jamie Harris, Ping Li, Chengfei Jiang, Hang Sun, and Haiming Cao. 2024. "An Integrative Transcriptome Subtraction Strategy to Identify Human lncRNAs That Specifically Play a Role in Activation of Human Hepatic Stellate Cells" Non-Coding RNA 10, no. 3: 34. https://doi.org/10.3390/ncrna10030034