Figure 3.
Reduction of PLIN2 binding to adiposomes by PtdIns
Adiposome preparation with dose of PtdIns (OD600 = 20) was aliquoted equally to 30 μl each and incubated with 0.5 μg SMT3-PLIN2-GFP per aliquot at 37°C for 5 min. After reisolation, washing and stained with LipidTOX Red (red), the PLIN2 (green)-coated adiposomes were visualized using confocal microscopy, and the colocalization PLIN2 (green) and adiposomes (red) were quantified by Pearson correlation analysis using ImageJ. The images were grouped as (a) SMT3-PLIN2-GFP, (b) LipidTOX Red, (c) merged signals, and (d) Pearson coefficient of correlation plot.
(A) Adiposomes were made with DOPC only (DOPC).
(B) Adiposomes were made with 99.6% DOPC and 0.4% PtdIns, in molar ratio (0.4% PtdIns).
(C) Adiposomes were made with 92.4% DOPC and 7.6% PtdIns, in molar ratio (7.6% PtdIns). Scale bar = 2 μm.