Fig. 2.

Evt-2 PH binds to PS. (A and B) His-tagged evt-2 PH (WT or K20E) was mixed with liposomes harboring the indicated negatively charged lipid (20% mol/mol of total lipids). After 15 min, the mixture was spun at 100,000 g for 30 min, and the resultant supernatant (S) and pellet (P) were subjected to SDS-PAGE. The gels were stained with Coomassie blue (A). The intensities of individual bands were quantitated with NIH ImageJ. The values of “P (bound)/S (free)” are shown in a bar graph (B). Data represent mean values ± SD of three independent experiments. PE, phosphatidylethanolamine; PC, phosphatidylcholine; PA, phosphatidic acid; PI, phosphatidylinositol. (C) Confocal images of wild-type yeast cells and cho1Δ cells expressing GFP-tagged 2 × PH (a tandem fusion of evt-2 PH) or GFP–Lact-C2. (Scale bars, 10 μm.) (D) COS-1 cells were transfected with GFP–Lact-C2, then fixed, and stained for GM130 or TfnR. Arrowheads indicate PM. (Scale bars, 10 μm.)