Figure 2. Neutral lipids accumulate in... | Rockefeller University Press

Figure 2.

$Figure 2. Neutral lipids accumulate in the ER in the absence of PAH1. (A) BODIPY, Nile red, and Oil red O staining of wild type (WT), pah1Δ, 4KO (dga1Δlro1Δare1Δare2Δ), and 5KO (pah1Δdga1Δlro1Δare1Δare2Δ) strains grown in SD medium. Images incorporate both brightfield and fluorescent channel for the indicated lipophilic dyes. Bar, 5 µm. (B) Inclusions in pah1Δ membranes seen with transmission electron microscopy. The indicated strains were cultured overnight in oleic acid medium before fixation. pah1Δ insets are higher magnifications of boxed areas and are shown to illustrate membrane inclusions. Note the membrane proliferation but no inclusions in the 5KO stain. LD, lipid droplet. (C) Droplet (LD) and membrane (M) levels of TAG and StE in the indicated strains. Postnuclear extracts from spheroplasts (grown in oleate) were fractionated by centrifugation. An identical percentage of the floating droplets and membrane pellets, normalized to total protein in the extracts, was subjected to TLC. Error bars indicate the range of values from two independent experiments.$

Neutral lipids accumulate in the ER in the absence of PAH1. (A) BODIPY, Nile red, and Oil red O staining of wild type (WT), pah1Δ, 4KO (dga1Δlro1Δare1Δare2Δ), and 5KO (pah1Δdga1Δlro1Δare1Δare2Δ) strains grown in SD medium. Images incorporate both brightfield and fluorescent channel for the indicated lipophilic dyes. Bar, 5 µm. (B) Inclusions in pah1Δ membranes seen with transmission electron microscopy. The indicated strains were cultured overnight in oleic acid medium before fixation. pah1Δ insets are higher magnifications of boxed areas and are shown to illustrate membrane inclusions. Note the membrane proliferation but no inclusions in the 5KO stain. LD, lipid droplet. (C) Droplet (LD) and membrane (M) levels of TAG and StE in the indicated strains. Postnuclear extracts from spheroplasts (grown in oleate) were fractionated by centrifugation. An identical percentage of the floating droplets and membrane pellets, normalized to total protein in the extracts, was subjected to TLC. Error bars indicate the range of values from two independent experiments.