Fig. 5. Effect of RGSZ1 expression on intracellular trafficking of VSVG-GFP. HeLa cells were transfected with an expression plasmid for VSVG-GFP with a control vector or a FLAG-RGSZ1 expression plasmid. The cells were incubated for 20 hours at 40°C, and then the temperature was changed to 32°C. At the indicated times the cells were fixed for immunofluorescence analysis. RGSZ1-expressing cells were identified by using the anti-FLAG antibody. The compartment to which VSVG-GFP had been transported at the indicated times after the temperature shift was determined. ER, G, FG and PM denote ER, Golgi, fragmented Golgi and plasma membrane, respectively. Values are expressed as a percentage of the total number of cells expressing both VSVG-GFP and RGSZ1. More than 100 cells were scored for each time point.