Localization of Fur4-GFP.

(A) KNP95 (Δura4 fur4-GFP) cells carrying pAU-gms1-RFP (ura4⁺) were cultured in EMM medium for 12 h at 30°C and observed using fluorescence microscopy. Arrows show co-localization of Fur4-GFP with Golgi marker and arrow heads show only localization of Fur4-GFP (top panel). KNP74 (fur4-GFP) cells were cultured in EMM medium for 12 h at 30°C and shifted to EMM containing 8μM FM4-64. These cells were cultured for 1 h and washed twice with EMM medium to visualize vacuoles and Fur4-GFP (bottom panel). (B) KNP74 (WT fur4-GFP), KNP95 (Δura4 fur4-GFP), and KNP83 (Δpub1 fur4-GFP) cells were grown in YES medium for 12 h. These cells were washed twice with sterile water and suspended in YES or YPD medium and incubated at 30°C for 1 h. These cells were observed using fluorescence microscopy. Bars: 10 μm.