Fig. 1. Cdc14p levels are cell-cycle regulated. (A) Structural comparison of the S. cerevisiae Cdc14p (ScCdc14p) phosphatase with the C. albicans Cdc14p homolog (CaCdc14p). White box, PTP motif; black box, putative NLS; hatched box, PEST sequence; black dots, Cdc28 phosphorylation sites. (B,C) Small G1 cells carrying Cdc14p-HA (JC94) were isolated by elutriation and released into (B) YPD at 30°C (yeast growth) or (C) YPD + 5% FCS at 37°C (hyphal growth). Samples were collected at intervals of 15 minutes after release and were assayed by western blotting for Cdc14p-HA levels. (D) Mitotic extracts corresponding to sample at 120 minutes from experiment B (- FCS) or sample at 135 minutes from experiment C (+ FCS) were treated with (+) or without (-) 
phosphatase ( PPTase) for 30 minutes. Samples were separated by SDS-PAGE and probed with anti-HA antibodies (12CA5). Anti-PSTAIRE antibody to detect Cdc28p was used as loading control. (E) Ratio of phosphorylated (Cdc14p-PO4) versus unphosphorylated (Cdc14p) forms of Cdc14p during yeast (black line) and hyphal growth (gray line). The ratio was calculated using Adobe Photoshop 7.0 software to quantify the intensity of the signals of the blots.