BODIPY staining and microscopy for evolved and engineered <i>KSR1</i> mutants.

<p>(A) Fluorescent microscopy images for cells from strains engineered to contain the <i>KSR1</i> LOH1 and to be completely homozygous for <i>KSR1</i> for either the A or B allele. Cells were stained with the lipid droplet dye BODIPY after growth in YPD alone (top), after exposure to 128 μg/mL FLC (middle), or after exposure to 128 μg/mL myriocin (MYO, bottom). (B) Quantification of the number of lipid droplets, lipid size, and integrated pixel intensity for fluorescence microscopy images, a subset of which are 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 are an average of at least 200 cells and error bars represent standard errors of the mean (SEM). (C) As in A, fluorescence microscopy images for the three additional evolved strains with LOH affecting <i>KSR1</i>. (D) Quantification of fluorescence microscopy, a subset of which is 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>