Fig 4. Lipid biosynthesis of 1Δ2Δ3tyΔ mutants assessed by metabolic labeling.

(A–C) WT or 1Δ2Δ3tyΔ cells incubated with or without 10 μg/ml Doxy for the indicated times in YPD (A) or YPD + 1.4 M sorbitol (B), were radiolabeled with [³H]-C16:0 for the indicated times at 30°C, the lipids were extracted, treated with NaOH as indicated, spotted and separated by TLC using solvent 1. (C) same as in (B) but additional 25 μM of cold C16:0 were added during labeling and lipids were separated by TLC with solvent 2. (D) cells grown with or without 10 μg/ml Doxy for 16 h in inositol-free SC + 1.4 M sorbitol were labeled with [³H]-myo-inositol for 2 h and the lipids from 10 OD₆₀₀ of each cell type were processed as in (C). NL = neutral lipids are free FAs, DAG, TAG, ergosterol esters and (acyl-)ceramides.