FIG. 2.

Phenotypic analysis of ncs1Δ::LEU2 cla4Δ strains. (A) Loss of NCS1 is lethal in the absence of CLA4. Strains SY3357 (wild type), SY3364 (ncs1Δ::LEU2 cla4Δ::TRP1 <pRS316ADE8CLA4>), SY3365 (ncs1Δ::LEU2 cla4Δ::TRP1 <YCpHIS3cla4-75>), and SY3360 carrying the cla4-75 allele (cla4Δ::TRP1 <YCpHIS3cla4-75>) were grown in synthetic medium at 25°C to an OD₆₀₀ of 1.0 to 2.0. A serial dilution (1/10) was performed starting with 10,000 cells. Cells were spotted onto prewarmed YEPD plates at 25 and 37°C and allowed to grow for 3 days. (B) Morphological examination of ncs1Δ::LEU2 cla4Δ::TRP1 strains. Strains SY3364, SY3365, and SY3360 (carrying YCpHIS3cla4-75) were grown in synthetic medium at 25°C and shifted to 37°C for 4 h.