Figure 1.

Liposome pull down assay. Liposomes with a fixed diameter of 0.2 μm were prepared using bovine brain lipid extract in HEPES-KCl buffer with 0.3 M sucrose, with (+) or without (−) exogenous cholesterol (20% w/w). Increasing amounts of liposomes (5, 10, 20 or 40 μL of a 2 mg/mL lipid suspension) were incubated with 50 μg of Der p 2, or glutathione-S-transferase (GST) for 30 minutes at 37 °C. The incubation mixture was then centrifuged at 30000 rpm, and the resulting supernatant (s) and pellet (p) were separated on a 12% SDS-PAGE gel. The gel was then stained with Coomasie Blue to visualize the amount of proteins bound to the liposome (p), or unbound (s). Full-length gels are presented in Supplementary Fig. S1.