Propranolol treatment caused the missorting of GFP-Ypt7p. Yeast cells expressing GFP-Ypt7p were grown in YPD and treated with buffer or propranolol for 12 h. Next, the vacuoles were pulse-labeled with FM4-64 for 1 h, washed with PBS, and chased with fresh YPD with or without propranolol for 3 h. After the chase, the cells were washed with PBS and prepared for observation by fluorescence microscopy. GFP-Ypt7p, FM4-64, and differential interference contrast (DIC) images were acquired separately. The GFP and FM4-64 images were merged using Photoshop.