Lipid droplets as ubiquitous fat storage organelles in C. elegans
1 Stowers Institute for Medical Research, 1000 E. 50th Street, Kansas City, MO 64110, USA
2 Deartment of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, KS 66160, USA
BMC Cell Biology 2010, 11:96 doi:10.1186/1471-2121-11-96Published: 8 December 2010
Additional file 1:
Dual labelling of BODIPY in live animals. In live wild-type animals, red BODIPY labelled GLO-1::GFP-encircled LROs with high intensity (arrowheads). It also labelled non-GLO-1::GFP-encircled structures with low intensity (arrows). Images were single confocal slices.
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Additional file 2:
Isolated lipid droplets free of LRO marker protein GLO-1::GFP. Lipid droplet fraction and cytoplasm fraction were prepared from stage L4 dhs-28;glo-1::gfp animals. Triplicates. Fractions were probed for the presence of GLO-1::GFP protein using a GFP antibody. GLO-1::GFP (predicted MW, ~50 kD, arrow) was detected in cytoplasm fractions but not in lipid droplet fractions. Asterisk denotes a non-specific protein recognized by the GFP antibody that is present in wild-type control (WT).
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Additional file 3:
Distinct emission spectra of lipid droplets and membrane pellets isolated from dhs-28 animals that were vital-labelled by Nile Red. (A) In stage L4 dhs-28 animals that were grown and stained on 10-cm NGM/OP50/Nile Red plates, LROs displayed Spec-1 emission spectrum (pseudo-colored in red) and lipid droplets displayed Spec-2 (pseudo-colored in green). Similarly, membrane pellets (B) and lipid droplets (C) isolated biochemically from these dhs-28 animals displayed Spec-1 and Spec-2 respectively. (D) Emission spectrum profiles of Spec-1s (red line) and Spec-2s (green line) of intact dhs-28 animals and Spec-1s (dashed red line) and Spec-2s (dashed green line) of isolated membrane pellets and lipid droplets. Intensity values were normalized.
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Additional file 4:
The same fluorescence emission spectrum of BODIPY-labelled LROs and lipid droplets. In live dhs-28 mutants, red BODIPY labelled both lipid droplets and LROs (marked by GLO-1::GFP). However, red BODIPY displayed the same emission spectrum (colored in gold) in lipid droplets and LROs. LD, lipid droplet. Images were single confocal slices.
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