Figure 4.

Hrr25p promotes Crz1p cytosolic localization. (A) YPH499 cells expressing 3xGFP-Crz1p (pLMB127) and either pCu423CUP1 or pKK194 (2µHRR25) were induced for 3 h with 500 µM CuSO₄. Then 200 mM CaCl₂ was added, and cells were examined at the indicated times after CaCl₂ addition. For each time point, 150-200 cells were counted, and the graph represents the average of two independent experiments. Gray bars represent cells with exclusively nuclear (N) GFP-Crz1p; white bars represent cells with intermediate nuclear/cytosolic localization (N/C); black bars represent cells with cytosolic localization (C). (B) Hrr25p depletion promotes Crz1p localization in low concentrations of Ca²⁺. KKY387 cells expressing 3xGFP-Crz1p were grown for 3 h with or without glucose. Then 50 mM CaCl₂ was added where indicated, and cells were examined within 5 min of CaCl₂ addition.