FIG. 6.

Localization of GFP-Ypt10p in yeast and HeLa cells. (A) GFP-Ypt10p was expressed in yeast cells, live 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. A merge of the two images is shown. (i) GFP-Ypt10p expressed from endogenous promoter in ypt10Δ cells; (ii) GFP-Ypt10p expressed with P_CUP1 (image shows basal expression of gene in absence of Cu³⁺ addition to media); (iii) live-cell imaging of GFP-Ypt10p in cells labeled with FM4-64 for 15 min followed by a 45-min washout to identify vacuolar membranes. (B) GFP-Ypt10p was expressed in HeLa cells, fixed and permeabilized, and labeled with TRITC-phalloidin to visualize the actin cytoskeleton. Cells were viewed by epifluorescence microscopy, and images were subjected to double-blind deconvolution. A single transverse slice though the cell is shown. GFP and TRITC-phalloidin are shown both individually and as a merged image. The panels below show GFP-Ypt10p in the periphery of the cell (i) and in conjunction with markers of the Golgi apparatus: GM130 (ii), GFP early endosome EEA1 (iii), and the recycling endosome for Rab11 (iv).For panels ii, iii, and iv, GFP and antibody markers are shown in separate images.