Title Page
Contents
Preface 6
Abstract 7
Graphical Abstract 9
국문초록 10
List of Abbreviations 15
1. Introduction 16
2. Materials and Methods 19
2.1. Mice 19
2.2. Transient transfection 19
2.3. Tamoxifen treatment 20
2.4. Real-time quantitative polymerase chain reaction (qPCR) 20
2.5. Immunoblotting analysis 20
2.6. Antibodies 20
2.7. Cell cultures 21
2.8. Retroviral transduction 22
2.9. Cytosolic and nuclear fractionation 23
2.10. Primary hepatic stellate cell isolation 23
2.11. Adenoviral infection of organoids 23
2.12. Conditioned media for LX-2 and primary hepatic stellate cell activation 23
2.13. Histologic analysis 24
2.14. Statistical analysis 25
3. Results 26
3.1. Duct-specific TAZ depletion inhibited liver fibrosis after a DDC diet 26
3.2. CD133-KO organoids decreased TAZ levels and secretion of fibrosis inducers 33
3.3. CD133 and TAZ levels were increased in bile ducts after a DDC diet 37
3.4. CD133-KO mice exhibited decreased fibrosis after a DDC diet 40
3.5. CD133 stimulated Src to stabilize TAZ 43
4. Discussion 47
REFERENCES 50
Supporting Information 61
Figure 1. Ductal WW domain-containing transcription regulator 1 (WWTR1)/transcriptional co-activator with PDZ-binding motif (TAZ)-specific... 30
Figure 2. Depletion of TAZ in cholangiocyte organoids decreased the expression levels of fibrotic markers. 32
Figure 3. Depletion of prominin 1 (PROM1/CD133) in cholangiocyte organoids decreased the TAZ levels. 36
Figure 4. DDC diet induced CD133 expression along with increase in TAZ levels. 39
Figure 5. CD133-KO mice showed decreased fibrosis after a DDC diet. 42
Figure 6. The CD133-Src-TAZ axis was responsible for the expression of fibrotic marker genes. 46