Abstract
Imaging host–pathogen interactions in real time can provide significant insight into dynamic processes and provide information about time and space of their occurences. Here, we present detailed experimental instructions on how to image the membrane penetration process of the non-enveloped adenovirus in real time. The system is based on a cell line stably expressing the lectin galectin-3 fused to a fluorophore. Membrane-lytic events during adenovirus cell entry can be monitored by the recruitment of galectin-3 to galactose-containing membrane glycoproteins on the exo-surface of ruptured membranes. The simultaneous use of fluorescently labeled adenoviral capsids allows to image the events in unmatched temporal resolution.
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Acknowledgements
Part of this work was supported by Equipe FRM 2011 Projet DEQ 20110421299 (H.W.). C.W. acknowledges funding from the NIH (AI082430) and American Heart Association (2261306). A.M.B. acknowledges support from the NIH (AI007508). We acknowledge the Bordeaux imaging centre (BIC) for help in setting up the live cell imaging acquisition. H.W. is an INSERM fellow.
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Martinez, R., Burrage, A.M., Wiethoff, C.M., Wodrich, H. (2013). High Temporal Resolution Imaging Reveals Endosomal Membrane Penetration and Escape of Adenoviruses in Real Time. In: Bailer, S., Lieber, D. (eds) Virus-Host Interactions. Methods in Molecular Biology, vol 1064. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-601-6_15
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DOI: https://doi.org/10.1007/978-1-62703-601-6_15
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-600-9
Online ISBN: 978-1-62703-601-6
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