Vacuolar phenotypes for evolved and engineered <i>KSR1</i> LOH mutants.

<p>(A) Fluorescent microscopy (top rows) and DIC (subsequent rows) images for strains engineered to contain the <i>KSR1</i> LOH1 or to be completely homozygous for <i>KSR1</i> for either the A or B alleles. Cells were stained with the vacuolar dye FM4-64 after growth in YPD without drug (top) or after exposure to 128 μg/mL FLC (bottom). (B) Quantification of vacuolar area and integrated pixel intensity for fluorescence microscopy images shown in panel A, (see <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1012497#sec011" target="_blank">Methods</a>). Points show an average of at least 200 cells and error bars represent standard errors of the mean (SEM). (C) As in A, fluorescence microscopy images of the additional three evolved strains with LOH affecting <i>KSR1</i>. (D) Quantification of fluorescence microscopy shown in panel (C). (B and D) Asterisks denote significant differences, using nonparametric t-tests, * P < 0.05, ** P < 0.01.</p> <p>(PDF)</p>