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. 2013 Apr 15;24(8):1196–1207. doi: 10.1091/mbc.E12-12-0852

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© 2013 Boulant et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 8: — Endosomal association of MRV virion and ISVP particles after cell entry. Cells transiently expressing Rab5-GFP or Rab7-GFP for 16 h were inoculated with fluorescent virus particles, and Z-series images were acquired from individual cells at different times postinoculation by 4D spinning-disk confocal microscopy, as described in Materials and Methods. (A) Maximum-intensity projection images of MDCK cells expressing early endosome marker Rab5-GFP (left panels) or late endosome marker Rab7-GFP (right panels) at 30 min postinoculation with both AF563-labeled virions (red) and AF647-labeled ISVPs (blue). White boxes correspond to the magnified insets. (B) Quantification of the fraction of virions and ISVPs (same particles as shown in A) colocalizing with Rab5-GFP (left panel) and Rab7-GFP (right panel). The number of virions or ISVPs in these endosomal compartments at different times postinoculation was normalized to the total number of virions or ISVPs in those cells at each respective time. The histogram represents ± SD for 10 cells analyzed. (C) Similar experiment as described in (B), but this time inoculating the cells with AF647-labeled ISVPs (blue) and AF563-labeled μ1(N42A)-ISVPs (magenta). The histogram represents the mean ± SD for 10 cells analyzed.