(A) Quantification of SEs in wild type and nvj1Δ cells collected at log phase, after 4 hr of AGR, or after a 4 hr co-treatment of AGR in the presence of 20 μg/mL lovastatin. Neutral lipids were extracted, separated by TLC, and visualized using Cu(II) Sulfate spray and charring. Quantification performed by densitometry. (Brown-Forsyth and Welch ANOVA. N = 6. **p-value<0.01) (B) Quantification of free sterols using the same strains and methodology as in (A). (Brown-Forsyth and Welch ANOVA. N = 6. ***p-value<0.001). (C) Scintillation counting quantification of radiolabeled HMG-CoA* produced after a 15 min pulse with 14C-acetate in intact WT or nvj1Δ cells grown for 4 hr in AGR. After quenching the radio-labeling reaction, soluble metabolites were isolated and endogenous HMG-CoA was converted into mevalonate using an in vitro HMGCR reaction. Mevalonate was subsequently separated and isolated by TLC, visualized by autoradiography, and quantified by scintillation counting. (Brown-Forsyth and Welch ANOVA. N = 6. **p-value<0.01). (D–G) Quantifications of radiolabeled DAG and mevalonate-derived lipids. Radio-labeling was performed as described in (C). Lipids were extracted and separated by TLC and visualized by autoradiography. Squalene and SE bands were quantified by scintillation counting. Ergosterol and DAG bands were quantified by densitometry. (Brown-Forsyth and Welch ANOVA. N = 6. ***p-value<0.001). (H) Growth curves of WT and nvj1Δ cells. Cells were treated with AGR for 10 hr in the absence or presence of 10 μg/mL mevalonate, and subsequently diluted to OD600 = 0.1 in SC media containing 2% glucose lacking mevalonate. OD600 measurements were taken every hour following dilution in fresh glucose-containing media. (I) Resumption probability graph depicting probability of cells producing a daughter bud as a function of time following an SC to SC+glucose transition (N > 85 cells).