Fig. 4.

ACA12 complements the phenotype of Ca²⁺-ATPase deficient yeast K616 strain. Precultures of K616 transformed with the specified constructs grown in synthetic complete medium lacking uracil (SC-URA), 2% (w/v) glucose, 10 mM CaCl₂ were pelletted, washed and diluted with water. Five µl of cells at A₆₀₀ = 1.0 - or serial 1:3 dilutions - were spotted on SC-URA, 2% (w/v) galactose, 1% (w/v) raffinose, supplemented with 10 mM CaCl₂ or 5 mM EGTA. All media were supplemented with 50 mM succinic acid/Tris (pH 5.5), 0.7% (w/v) yeast nitrogen base. Plates were incubated for 8 days at 20 °C. Results are from one experiment representative of three.