Data associated with Fig. 2 . Cells lacking Nem1, Spo7, Pah1, or Fld1 accumulate neutral lipids in the ER and have aberrant droplets. (A) Diagram depicting the regulation of triglyceride synthesis. The Nem1-Spo7 complex dephosphorylates and thereby activates the phosphatidate phosphatase Pah1, resulting in formation of DAG from phosphatidic acid (PA). DAG then serves as a substrate for the TAG-synthesizing enzymes Dga1 and Lro1. (B) BODIPY accumulates in the ER of nem1Δ, spo7Δ, and pah1Δ cells. Yeast cells of the indicated genotype expressing genomic Erg6-mCherry were stained with BODIPY. White arrowheads denote BODIPY accumulation in the ER membrane. Scale bar, 5 µm. (C) Fld1-deleted cells accumulate numerous small and a few supersized LDs. fld1Δ cells expressing genomic Erg6-mCherry were stained with BODIPY. White arrows denote colocalization between BODIPY and Erg6-mCherry. Yellow arrows denote BODIPY-stained supersized puntae that do not colocalize with Erg6-mCherry. Two examples are shown (I, II). Scale bar, 5 µm. (D) WT cells do not show BODIPY accumulation in the ER. BODIPY staining of WT cells expressing genomic Erg6-mCherry. White arrows denote colocalization between BODIPY and Erg6-mCherry. Scale bar, 5 µm. (E) Pah1-7P rescues the LD biogenesis defect of nem1Δ, spo7Δ, and pah1Δ mutant cells. BODIPY staining of nem1Δ, spo7Δ, and pah1Δ mutants expressing Erg6-mCherry and Pah1-7P. White arrows denote colocalization between BODIPY and Erg6-mCherry. Scale bar, 5 µm.