Biology Reference
In-Depth Information
CHAPTER
10
Gregory Camus
1
, Dorothee A. Vogt
1
, Andrew S. Kondratowicz, and Melanie Ott
Gladstone Institute of Virology and Immunology, University of California, San Francisco,
Owens Street, San Francisco, CA, USA
Lipid Droplets and Viral
Infections
CHAPTER OUTLINE
Introduction ............................................................................................................ 168
Interactions Between Viruses and LDs ..................................................... 169
Viruses that Localize to LDs ................................................................... 171
Viruses that use the Energy Stored in LDs................................................ 172
Viruses that Induce LD Rearrangement.................................................... 172
Viruses that use LDs as a protein-storage Depot ....................................... 173
10.1 Materials........................................................................................................173
10.1.1 Tools for Analysis of Viral Infection .............................................. 173
10.1.2 Tools for Analysis of LDs ............................................................. 174
10.1.3 Other Commercial Reagents and Drugs......................................... 174
10.1.4 Other Buffers and Reagents......................................................... 174
10.2 Methods .........................................................................................................175
10.2.1 Biochemical Cell Fractionation .................................................... 175
10.2.1.1 LD Isolation ....................................................................... 175
10.2.1.2 Cell Fractionation............................................................... 175
10.2.1.3 Membrane Flotation Assay ................................................. 176
10.2.2 Immunomicroscopy .................................................................... 177
10.2.2.1 Fixation and Labeling......................................................... 177
10.2.2.2 HCS LipidTOX Neutral Lipid Stain....................................... 177
10.2.2.3 Bodipy .............................................................................. 177
10.2.2.4 Oil-Red-O .......................................................................... 177
10.2.2.5 Quantification .................................................................... 178
10.2.3 Lipid Measurements ................................................................... 178
10.2.3.1 Triglycerides Extraction and Enzymatic Quantification ......... 179
10.2.3.2 Triglyceride Extraction from Cell Culture ............................. 179
10.2.3.3 Triglyceride Enzymatic Quantification ................................. 179
1
These authors contributed equally to this work.
