Expression of Torpedo and Arabidopsis CLC genes in gef1/gef1 diploids. (A) Vector pRS1024 (13) was introduced into wild-type and gef1/gef1 diploids. Plasmids pRG120, pRG175, pRG176, and pRG178 were introduced into the gef1/gef1 diploid. Approximately 104 cells of the indicated strains were grown for 4 days in YPD (lane 1), SD buffered with 50 mM Mes Tris (pH 7; Sigma) (lanes 2 and 3), and SGE buffered with 50 mM Mes Tris (pH 7) (lanes4 and 5). CuSO4 (Sigma) at 0.1 mM was added to lanes 3 and 5. Diploids were used because they gave a better discrimination of phenotypes. (B) Approximately 105 cells of the indicated strains were grown for 3 days in YPD (lane 6), 7 days in YPD supplemented with 1.75 M NaCl (lane 7) (Sigma), 3 days in YPD supplemented with 0.45 M tetramethylammonium chloride (lane 8) (Sigma), 3 days in YPD supplemented with 2 mM MnCl2 (lane 9) (Sigma), and 3 days in YPD supplemented with 2 mM MnSO4 (lane 10) (Sigma).