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Fig. 10. ARV1 deletion mutants exhibited endocytosis defects and fragmented vacuoles. (A) Lucifer yellow (LY) uptake and delivery to the vacuole was observed in wild-type (W.T.; CBY858), erg2{Delta} (CBY678) and arv1{Delta} (CBY994) cells grown at 30°C. In the wild-type strain, brightly stained vacuoles were observed in 92% of cells (154 out of 167 cells counted). In erg2{Delta} cells, only 2% of cells efficiently internalized lucifer yellow (1/50), and 0% of arv1{Delta} cells exhibited brightly staining vacuole or vacuolar fragments (0/72). (B) Wild-type (CBY858), las17{Delta} (CBY1024) and arv1{Delta} (CBY994) were treated with a short pulse of FM4-64 and then chased with fresh medium (FM4-64 P/C). Cells cultured at 30°C were viewed by fluorescence microscopy 30 minutes after the FM4-64 pulse/chase (P/C) and photographs were taken with equal exposures. (C) Wild-type (CBY858) and arv1{Delta} (CBY994) cells were cultured at 30°C in the presence of FM4-64 for 4 hours to label all vacuoles and vacuolar fragments uniformly (FM4-64). In arv1{Delta} cells, vacuolar fragmentation was evident in the proliferation of FM4-64 staining vacuole-derived rings.