Figure 6.

Negative Contrast Immunographs of High-Density Sucrose Fractions.

Arabidopsis P100 membranes were centrifuged under isopycnic conditions in a 15 to 55% (w/w) linear sucrose gradient. High-density fractions (47 to 50%) corresponding to the strong VPS35 signals in protein gel blots at this density (Figures 5A and 5B) were negatively stained. (A) is an overview, (B) is a high-magnification detail, and (C) and (D) show putative retromer vesicles positively stained with VPS35 and VPS26 immunogold, respectively. The arrows point to clathrin-coated vesicles; the arrowheads label putative retromer-coated vesicles. Bars = 100 nm.