Fig. 2. Confocal microscopy of VSVG-ts045-GFP protein transport during poliovirus infection. Huh7 cells were transiently transfected with VSVG-ts045-GFP for 16 hours at 40°C. (A) Uninfected cells were shifted to 32°C for 0, 10 or 20 minutes as indicated, fixed and visualized for GFP fluorescence by confocal microscopy. (B) Transfected cells were infected at an MOI of 30 PFU/cell, incubated at 40°C for 3 hours and shifted to 32°C for 0, 10 minutes or 20 minutes as indicated, fixed and visualized for GFP fluorescence by confocal microscopy. For each panel, an image was selected to represent most abundant class of expression pattern for that condition and time point. Arrows identify VSVG-ts045-GFP puncta and arrowheads identify Golgi-like patterns. (C) The numbers of cells with ER-like, ER with puncta, and Golgi-like patterns were determined for both infected and uninfected cells at 0, 5, 10 and 20 minutes after temperature shift. Three cover slips per time point were evaluated and the percentage of cells with each pattern category was determined. At least 50 cells per coverslip were evaluated. For each type of pattern, the percentage of cells is plotted as a function of time for uninfected () and infected () cells.