LD assembly causes phospholipid-packing defects in seipin complex mutants. (A) Localization of GFP-Pct1 and GFP-Spo20^51–91 in cells with (wt and ldb16Δ) and without LDs (are1Δ are2Δ dga1Δ lro1Δ and are1Δ are2Δ dga1Δ lro1Δ ldb16Δ). Cells were grown in SC media up to early stationary phase. LDs were stained with the neutral lipid dye MDH. Bar, 5 µm. (B) Localization of GFP-Pct1 and GFP-Kes1 at the indicated time points upon induction of LD formation. LDs were induced by expression of the triglyceride-synthesizing enzyme Dga1 from a regulatable promoter in are1Δ are2Δ lro1Δ (LD^Switch) or are1Δ are2Δ lro1Δ ldb16Δ (ldb16Δ LD^Switch) cells. LDs were stained with the neutral lipid dye MDH. Bar, 5 µm. (C) Kinetics of LD formation and appearance of GFP-Pct1 (left) or GFP-Kes1 (right) foci in ldb16Δ LD^Switch cells. Cells were grown in minimal media supplemented with 75 µM inositol until early stationary phase. The percentages of ldb16Δ LD^Switch cells displaying LDs (black line) or foci of the indicated GFP-tagged protein (gray line). Expression of Dga1 was induced at time zero. The mean of two independent experiments is displayed; error bars represent SD. At least 100 cells/time point were analyzed in each experiment.