Supporting Online Material

Repulsion of Superinfecting Virions: A Mechanism for Rapid Virus Spread
Virginie Doceul, Michael Hollinshead, Lonneke van der Linden, Geoffrey L. Smith

Supporting Online Material

This supplement contains:
Materials and Methods
Figs. S1 to S6

This file is in Adobe Acrobat PDF format.

Other Supporting Online Material for this manuscript includes the following:

Movie s1
Movie showing VACV plaque formation. BSC-1 cells were infected with VACV strain WR at low moi and the formation of a plaque was recorded by phase microscopy every h for 16 h after a small plaque first became visible. Note that the motility of infected cells is restricted to within the area showing cytopathic effect (cpe). Virus-induced cell motility is therefore not increasing the rate of spread across the cell monolayer.

Movie s2
Movie showing plaque formation by a VACV strain vEGFPA5L. This virus expresses EGFP fused to the A5 core protein late during infection. BSC-1 cells were infected with vEGFPA5L at low moi and the progression of infection on one side of a plaque was recorded by phase microscopy every h for 16 h.

Movie s3
Movie as for movie 2 except that the growth of the vEGFPA5L plaque was visualized by the EGFP fluorescence.

Movie s4
Movie as for movies 2 and 3 showing merge of phase and fluorescence images. Note that the spread of virus-induced cpe (an early event) precedes that of expression of EGFPA5L, which is made only late during infection.

Movie s5
Movie showing the spread of the edge of a plaque formed by vEGFPA5L at higher rmagnification to show individual virus particles (green dots). Cells were visualised by confocal microscopy using EGFP flourescence. Note cells must express EGFP prior to production of new virus particles and yet there are numerous virus particles on cells distal to those cells expressing EGFP.

Movie s6
Movie showing the formation of actin tails (red) in BSC-1 cells expressing cherryFP-actin that have been infected with vEGFPA5L (green). The movie shows a single focal plane at the periphery of an infected cell where actin tails are formed following the transport of virus particles to the cell periphery (duration, 5 min).

Movie s7
Movie showing green virus particles moving on the tip of red actin tails on a cell expressing cherry-actin and lacking green virus factories. Note that a virus-tipped red actin tail produced by this cell induces the formation of another actin tail after recontacting the cell surface.

To view these movies, download a QuickTime viewer.