Proteomics and fluorescence imaging indicates selective remodeling of LD proteome during AGR. (A) Cartoon schematic of step-wise LD isolation. (B) Western blot of whole cell lysate (WCL), and fractions of LD isolation protocol as in A. Pma1, plasma membrane marker; Por1, mitochondria marker; Pln1, LD marker; Tubulin, cytoplasmic marker. (C) Heatmap of log₂-fold abundance changes of annotated LD proteins from LC-MS/MS proteomics on either isolated LD samples (top) or whole-cell lysates (bottom). Heat map shows the relative log₂ change of 4 h AGR over log-phase samples. (D–K) Yeast with GFP or mNg-tagged LD proteins with MDH LD stain in log-phase and 4 h AGR. Each protein has quantification of log₂ fold abundance changes from LD fraction (LD) and whole-cell (WC) proteomics as in C, as well as the relative M1 Manders coefficient from imaging. (L) Yeast expressing GFP-Livedrop and imaged in either log-phase or 4-h AGR conditions, with corresponding M1 Manders coefficient quantified. (M) Snx4-mNg yeast with LD MDH stain in log and 4-h AGR yeast. Insets are of LDs and Snx4-mNg signal. Calculated log₂ fold abundance changes for LD, WC, and M1. ***, P < 0.001. All scale bars, 5 μm. Source data are available for this figure: SourceData F5.