FIG. 7.

Localization of GFP-Ypt11p in yeast and HeLa cells. (A) GFP-Ypt11p was expressed in yeast cells, and live cells were stained with Hoechst 33258. Cells were viewed by epifluorescence microscopy, and fluorescence images were subjected to double-blind deconvolution. The GFP fluorescence signal is presented with a differential interference contrast (DIC) image of the same cells, and a merge of the three images is shown. (B) GFP-Ypt11p was expressed in yeast cells, fixed, and stained with Hoechst 33258. Cells were analyzed by immunofluorescence microscopy using anti-Pdi1p antibodies. The GFP and immunofluorescence signal is presented with a differential interference contrast (DIC) image of the same cells. (C) GFP-Ypt11p was expressed in BHK cells and viewed by epifluorescence microscopy, and fluorescence images were subjected to double-blind deconvolution. Both a maximum projection and a single slice through the cell are shown. (D) GFP-Ypt11p was expressed in BHK cells and live cells incubated with ER-Tracker Blue-White DPX. Cells were fixed and viewed by epifluorescence microscopy. The GFP and ER-Tracker DPX signals are shown individually, and in a merged image with ER-Tracker DPX false-colored red to show colocalization with GFP. (E) GFP-Ypt11p was expressed in HeLa cells, and fixed cells were viewed by confocal microscopy. The GFP fluorescence signal was compared to that of an antibody marker for the endoplasmic reticulum, PDI. Images show the periphery of the cell both individually and as a merge of the two fluorescence images. (F) GFP-Ypt11p was expressed in BHK cells, and fixed cells were viewed by confocal microscopy. The GFP fluorescence signal was compared to that of an antibody marker for the ERGIC, KDEL receptor (KDEL-R). GFP and the KDEL-R are shown both individually and as a merged image. In the lower panels, the cells were incubated for 3 h at 15°C before fixation.