Journal: Virology

Article Title: Direct evidence for intracellular anterograde co-transport of M-PMV Gag and Env on microtubules

doi: 10.1016/j.virol.2013.11.006

Figure Lengend Snippet: Nocodazole treatment leads to a transient decrease in linear movement of intracellular capsids. CMMT cells were co-transfected with pSARM–GagGFP–M100A and pTML-Env–mCherry and visualized in real time with a Deltavision deconvolution microscope. Live imaging of untreated cells and those treated with nocodazole for 2 h and 4 h was performed. Videos were recorded over a 2 min time period, with each video having an identical frame rate of five frames per second.(A) Illustration of tracks for Gag–GFP (green), Env–mCherry (red) and co-localized Gag–Env particles (yellow) in untreated and nocodazole-treated cells. Tracks shown are for particles moving towards or near the plasma membrane, with arrow heads indicating direction of movement. Data shown is representative of at least five independent imaging experiments. Bars=15 μm. See accompanying Movies S2–S4 for real time visualization of particle tracking. (B) Maximum velocity measurements of Gag–GFP particles (n ≥ 15), Env– mCherry particles (n ≥ 10) and co-localized Gag–Env particles (n ≥ 23) in untreated cells and nocodazole-treated cells after 2 h and 4 h of drug treatment. A minimum of 5 cells was analyzed at each time point. Bars indicate median values. Statistical significance is denoted by p value or ns (not significant).

Article Snippet: Cells were visualized using a 60 × oil objective on a Deltavision deconvolution fluorescence microscope (Applied Precision Inc., Issaquah, WA).

Techniques: Transfection, Microscopy, Imaging, Clinical Proteomics, Membrane