Supplemental information
PhD thesis: “Visualization of the intracellular
trafficking of Adeno-associated virus type 2”
Kerstin Lux
1. Movies:
Movie 1 chapter 2.1.4: HeLa dsRed Nuc cells infected with
100%-GFP-VP2-AAV (approx. 106 capsids/cell).
Movie generated by conventional time laps
microscopy:
27 images were taken with a time laps of 30
seconds.
Some GFP-tagged AAV virions seem to touch the
cell membrane multiples times. Most virions stack to the membrane, but some GFP
tagged virions seem to move inside the cytoplasm. These observations confirm
previous observations made by SVT.
Movies 2 and 3: HeLa cells infected with Cy5 labeled AAV
(approx. 300 capsids/cell).
Real time movies generated by SVT measurements:
directed movements of Cy5 labeled AAV inside HeLa cells.
Images were taken every 40 milliseconds. Movie
2 contains 200 and movie 3 contains 110 frames.
Movie 4: PtK2 cells infected with
100%-GFP-VP2-AAV (approx. 106 capsids/cell).
Movie generated by conventional time laps
microscopy:
Images were taken with a time laps of 2
seconds, 100 images were taken.
GFP tagged AAV particles can be observed
performing directed movements. Furthermore, many particles perform small,
oscillatory movements without making large displacements.
2. PDF files of publications:
Lux et al
2005.pdf: “Green fluorescent protein-tagged adeno-associated virus
particles allow the study of cytosolic and nuclear trafficking”
Perabo and Endell et al 2006.pdf : “Combinatorial engineering
of a gene therapy vector: directed evolution of adeno-associated virus”
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