Figure 2.

GFP-Bud6 localization during the cell cycle in wild-type haploids or diploids. (A) Selected frames from a 153-min time-lapse series showing representative stages of GFP-Bud6 localization in wild-type haploids (MY15DBG) during the cell cycle. Bud6 initially localized to the prebud site. Association to the neck area occurred at ∼15 min after bud emergence (left cell, large arrowhead). Appearance of a second ring at the daughter face of the neck occurred at ∼110 min (small arrowhead). Numbers indicate time in minutes relative to bud emergence for the cell on the left. Precise timing of label association to the cell on the right could not be determined because the growing bud on the left pushed the cell neck off focus. Bar, 2 μm. (B) Selected frames from a 169-min time-lapse series showing initial association of GFP-Bud6 to the bud tip followed by faint labeling of the neck region (arrowhead) in a wild-type diploid (MY15DdBG). Label on the bud surface remained concentrated at the distal portion of the bud until 30 min before cytokinesis. Numbers indicate time relative to bud emergence. Bar, 2 μm. (C) Plot showing linescans through the polarity axis reflecting the relative association of GFP-Bud6 to the neck and bud surface in a haploid or diploid wild-type cell at a fixed mother/daughter size ratio corresponding to ∼30 min after bud emergence.