Figure S5.

Hydroxyl-containing residues in yeast Ldb16 are required for correct LD morphology in TAG-only and SE-only cells. Related to Fig. 7. (A) Analysis of LD morphology in cells with the indicated genotype by fluorescence microscopy. Cells were cultured in inositol free media at 30°C to late stationary phase, and LDs were stained with BODIPY. Scale bars correspond to 5 μm. (B) Quantification of LD size in cells shown in A. A minimum of 100 LDs per genotype and two independent biological replicates were quantified. Bars and whiskers indicate mean with interquartile range. (C) Analysis of LD morphology in cells with indicated genotype upon plasmid borne overexpression of the NL enzymes Dga1 or Are1 which synthesize TAG or SE, respectively. Micrographs were take after 24 h addition of galactose to induce the expression of the NL synthesizing enzymes. LDs were visualized using the NL dye BODIPY 493/503. Scale bars correspond to 5 μm. (D) Analysis of LD biogenesis upon induction of SE synthesis by expression of the SE acyltransferase Are1. Plasmid-borne expression of Are1 was induced by addition of galactose (final concentration 2%) to cells with the indicated genotype grown in raffinose-containing medium. LDs were stained with BODIPY at indicated timepoints (in hours after galactose induction). Scale bars correspond to 5 μm.