Fig 3.

Pho85 inactivation leads to downregulation of Cln3. (A) Wild-type (wt) and pho85Δ cells were grown exponentially in YPD and then assessed for levels of Cln3-Myc (left), and wild-type cells were grown in synthetic complete medium with (+PO₄²⁻) or without (−PO₄²⁻) phosphate (right). Samples were taken after 6 h, and levels of Cln3-Myc were monitored. (B) Cells from the YAM67 strain were incubated with either 1-Na PP1 (a specific pho85-as inhibitor) or drug vehicle (dimethyl sulfoxide [DMSO]). Samples were taken at the indicated times, and Cln3-Myc was analyzed by immunoblotting. Data ± standard deviations from three independent experiments are shown. Cln3p, phosphorylated Cln3.