FIGURE 7.

Age1-GFP localization at the trans-Golgi and endosome. A, with galactose as the sole carbon source, cells were grown at 23 °C, diluted in fresh medium, and grown for an additional 16 h at 23 or 37 °C before visualization. The localization of Age1ΔN-GFP, expressed under control of the GAL1 promoter, was assessed in wild-type cells at 23 °C and 37 °C. Cells were visualized by fluorescence (left panels) and differential interference contrast microscopy (right panels). B, the colocalization of Age1ΔN-GFP and Age1-GFP, expressed under control of the GAL1 promoter, with RFP organelle markers was assessed in age2Δ gcs1-4 cells at 37 °C. Arrows indicate representative colocalization. C, the localization of Age1-GFP, expressed under control of the AGE1 promoter, was monitored in proliferating age2Δ gcs1-4 cells also carrying an empty vector or a high-copy SFH2 plasmid and incubated for 2 h at 37 °C. D, the localization of Age1ΔN-GFP and Age1-GFP, expressed under control of the GAL1 promoter, was analyzed at 37 °C in age2Δ gcs1-4 cells with the wild-type SPO14 gene or a spo14Δ deletion mutation.