Figure 4
From: Morphologically and Functionally Distinct Lipid Droplet Subpopulations
Separation and neutral lipid analysis of lipid droplet subpopulations from mouse liver.
LDs from mouse liver were fractionated by size using differential centrifugation (Materials and Methods). Four LD fractions were collected. (A) LD size of the subpopulations. LD fractions were viewed under light microscope (upper panel). The LDs were incubated with 1% Triton for 30 min to reduce aggregation. After removing Triton, the LDs were viewed and the images were acquired at 400-fold magnification. The LD size was measured by a Delsa Nano C particle analyzer. (B) Protein profile of the subpopulations. The proteins of LD subpopulations were extracted and analyzed by silver staining. (C) ER and mitochondrial markers in the subpopulations. Proteins from the subpopulations were analyzed by Western blotting with indicated antibodies (equal amount of protein per lane). (D,E) Neutral lipid analysis. Lipids of the subpopulations from mouse liver (panel D) and CHO K2 cells (panel E) were separated by TLC with a solvent of n-hexane-diethyl ether-acetic acid (80:20:1, v/v/v). TLC plate was then stained in iodine vapor after being dried at room temperature.
