FIG. 3.
Localization of ectopically expressed wild-type and mutant forms of Cdc5 in a diploid wild-type strain, 1788. To localize Cdc5, EGFP-Cdc5 fusion constructs were generated and expressed under the control of the GAL1 promoter for 4 h. Transformants expressing EGFP fusion constructs were stained with propidium iodide to visualize chromosomal DNA and examined by confocal microscopy. Arrows indicate rings at mother bud necks. Note that EGFP-Cdc5 forms a ring structure at the mother bud neck (NA panel). Vector, YCplac22-GAL1; NA, YCplac22-GAL1-HA-EGFP-cdc5N209A; WT, YCplac22-GAL1-HA-EGFP-Cdc5; FAA, YCplac22-GAL1-HA-EGFP-cdc5W517F/V518A/L530A. DIC, differential interference contrast; Cdc5, EGFP-Cdc5 expression; PI, propidium iodide staining of nuclei; Cdc5 + PI, EGFP-Cdc5 and propidium iodide images superimposed. Bar, 5 μm.