Elucidating the role of cathepsin B in the lifecycle of influenza A virus

.................................................................................................................................... ii Acknowledgements.................................................................................................................. iii Table of Figures ....................................................................................................................... vi Table of Appendices ............................................................................................................... vii Abbreviations......................................................................................................................... viii Chapter 1: Introduction ............................................................................................................. 1 1.1 Influenza A Virus.............................................................................................................1 1.1.1 IVA Structure............................................................................................................1 1.1.2 IVA Genetics ............................................................................................................4 1.1.3 IVA Lifecycle ...........................................................................................................5 1.1.4 Hemagglutinin ..........................................................................................................9 1.2 Cathepsin B....................................................................................................................10 1.2.1 Background.............................................................................................................10 1.2.2 The role of Cathepsin B in viral lifecycles .............................................................11 1.2.3 Potential involvement of Cathepsin B in lipid homeostasis ...................................12 1.3 Rationale, hypothesis and objectives .............................................................................13 Chapter 2: Methods................................................................................................................. 14 2.1 Reagents .........................................................................................................................14 2.2 Cell Culture....................................................................................................................14 2.3 Viral Infections ..............................................................................................................15 2.4 Western Blot ..................................................................................................................15 2.5 RT-qPCR........................................................................................................................15 2.6 Fractionation ..................................................................................................................16 2.7 Cholesterol Assay ..........................................................................................................17 2.8 Protein Assay .................................................................................................................17 2.9 Immunofluorescence Microscopy..................................................................................17 2.10 Hemagglutination Assay..............................................................................................18 2.11 MTT Assay ..................................................................................................................18 2.12 Statistical Analysis.......................................................................................................19