Figure 1.

PLIN2 targets to lipid droplets and adiposomes

Lipid droplets or adiposomes were stained with LipidTOX Red (1:1,000, v/v) and endogenous PLIN2 or recombinant PLIN2 were labeled by GFP (green).

(A) PLIN2-GFP targets on surface of LDs. PLIN2–GFP was knocked in C2C12 cells, and the cells were treated with 100 μM oleate for 12 h and then imaged by Olympus FV1200 confocal microscope.

(B) PLIN2-GFP targets on lipid droplets isolated from C2C12 cells. The lipid droplets were isolated from C2C12 cells with the same treatment and imaged by Zeiss Image M2 microscope.

(C) Recombinant SMT3-PLIN2-GFP targets on adiposomes. The adiposomes (30 μl) prepared from DOPC and TAG were incubated with 5 μg SMT3-PLIN2-GFP at 37°C for 5 min. After reisolation, fluorescence images of adiopsomes were captured using a DeltaVision OMX (SIM) microscope. The images were grouped as (a) endogenous PLIN2-GFP or SMT3-PLIN2-GFP, (b) LDs or adiposomes both stained with LipidTOX Red, (c) merged signals. Scale bar = 2 μm.

(D) The diameter distribution analysis of intracellular LDs, isolated LDs (intracellular) and adiposomes in fluorescence images. The diameter of LD or adiposome was measured and analyzed using ImageJ to scan three images for each group.

See also Figures S1–S3 and Table S1.