Fig. 1. Transport of VSVG-ts045-GFP protein during poliovirus infection. HeLa cells were transfected with VSVG-ts045-GFP-expressing plasmid and incubated at 40°C overnight to accumulate VSVG-ts045-GFP protein in the ER. (A) Uninfected HeLa cells were either maintained at 40°C (A, left image) or shifted to 32°C for 30 minutes as indicated. The cells were subsequently fixed and mounted for fluorescence microscopy. (B) VSVG-ts045-GFP transfected cells were infected with poliovirus at an MOI of 0.5 PFU/cell at 40°C for 3.5 hours then shifted to 32°C for 30 minutes Cells were then fixed and analyzed for GFP fluorescence or the presence of poliovirus 2C protein by indirect immunofluorescence as indicated to distinguish between uninfected cells and infected cells in the same image. Arrowheads indicate Golgi complexes, arrows indicate ERGIC puncta.