FIG. 2.

Cla4 requires vacuole inheritance for vacuole-associated punctate structure localization and remains on the daughter vacuole until late M phase. (A) Asynchronous cultures of FM 4-64-stained Cla4-GFP (Y_CH3250) were examined by fluorescence microscopy and each cell was categorized as small, medium, or large budded (>200 cells were counted for each bud size). Each cell was examined for both vacuole morphology (green) and Cla4-GFP localization to a punctate spot (red), and all cells fell into one of the categories shown. The percentages of small, medium, or large budded cells with the indicated vacuole morphology and punctate Cla4 localization are listed. (B) CLA4-GFP (Y_CH3250) localization was visualized in cells of various bud sizes and in cells arrested in S phase with hydroxyurea (HU), at G₂/M with nocodazole (NZ), or at mitotic exit by arresting cdc15-2 (Y_CH1028). Values indicate the percentages of cells with the displayed Cla4-GFP localization. (C) Cla4-GFP was visualized in vac8 (Y_CH1665) and myo2-2 (Y_CH1752) cells. Arrowheads indicate Cla4-GFP localization at the cortex. (D) Cla4-GFP and FM 4-64-stained vacuole localization was quantified in small, medium, and large budded wild-type (Y_CH3250), vac8 (Y_CH1665), and myo2-2 (Y_CH1752) cells (>200 cells were counted for each bud size).