Title Page

Contents

ABSTRACT 8

INTRODUCTION 9

MATERIALS AND METHODS 12

2.1. Materials 12

2.2. Cell culture 13

2.3. Superoxide measurement 13

2.4. Transfection of Plasmid DNA and si-RNA 13

2.5. Western blotting 14

2.6. Immunoprecipitation (IP) 14

2.7. Cell fractionation 15

2.8. Monolayer Wound Healing Assay 15

2.9. Chromatin immunoprecipitation (ChIP) and PCR 16

2.10. Immunofluorescent Staining 16

2.11. Quantitative Real-Time Polymerase Chain Reaction (qPCR) 17

2.12. Statistical analysis 17

RESULTS 18

3.1. LPS induces superoxide production through ROCK2 and p-p47phox. 18

3.2. LPS upregulates p-Tyr42 RhoA phospholipase D1 (PLD1) levels with forming a protein complex. 21

3.3. p-Tyr42 RhoA is involved in the EMT process and cell migration. 24

3.4. p-Tyr42 RhoA and PLD1 bind to the promoter of ZEB1, influencing cell migration 27

3.5. MYH9 is a PA-binding protein and forms a complex with p-Tyr42 RhoA and PLD1 32

DISCUSSION 38

CONCLUSION 42

REFERENCES 43

ABSTRACT IN KOREAN 47

APPENDIX 49

LIST OF ABBREVIATIONS 53

SUPPLEMENT PUBLICATIONS 54

Article 55

Figure 1. LPS increases superoxide generation via ROCK2 and p-p47phox. 19

Figure 2. LPS upregulates p-Tyr42 RhoA, phospholipase D1 (PLD1) levels via generating a protein complex. 23

Figure 3. EMT and cell migration are facilitated by p-Tyr42 RhoA. 26

Figure 4. The ZEB1 promoter is affected by the binding of p-Tyr42 RhoA and PLD1, leading to a modulation of cell migration 30

Figure 5. MYH9 is a PA-binding protein. 35