Phospholipids species in WT and Δopi3 cells. Cells were grown to log phase in SD‐URA, and shifted to starvation (SD‐N) without choline (left panel, −Choline), or supplemented with choline (1 mM) during nitrogen starvation only (Right panel, +Choline: SD‐N). Cells were harvested after 4 h of starvation and analyzed by shotgun lipidomics. Statistical analysis was done by ANOVA multiple comparisons test‐Sidak's (****P ≤ 0.0001, **P ≤ 0.005, *P ≤ 0.05, ns, not significant), error bars represent SEM of at least 3 independent experiments.
Acyl chain length and saturation of PC in WT and Δopi3 cells. Cells were grown and analyzed as in A.
Phospholipid composition of autophagic membranes isolated from Δypt7 and Δopi3Δypt7 cells expressing GFP‐Atg8. Cells were grown to log phase in SD‐URA, and shifted to starvation (SD‐N) without choline (left panel, −Choline), or supplemented with choline (1 mM) during nitrogen starvation only (right panel, +Choline: SD‐N). Cells were harvested after 3 h of starvation and autophagic membranes were isolated and analyzed by shotgun lipidomics as detailed in Materials and Methods. Statistical analysis was done by ANOVA multiple comparisons test‐Sidak's (****P ≤ 0.0001, **P ≤ 0.005, ns, not significant), error bars represent SEM of 3 independent experiments.
Acyl chain length and saturation of PC in autophagic membranes isolated from Δypt7 and Δopi3Δypt7 cells expressing GFP‐Atg8. Cells were grown and analyzed as in C.