Figure 5

Purification of PGL34-YFP by flotation centrifugation. A. Diagram illustrating the purification procedure. Suc., sucrose concentration. B. Western blot analysis of fractions from the density gradient. After ultracentrifugation, 0.5 ml fractions were collected starting from the top of the gradient. Proteins contained in 400 μl of fractions 1–7, 200 μl of fractions 9–15, 100 μl of fractions 17–19 or 50 μl of fraction 21 were separated by SDS-PAGE, transferred to a nitrocellulose membrane and stained with amidoblack (upper panel). Silver stained plastoglobule proteins prepared from wild type plants (wt) are shown. Antibodies against GFP, PGL35, TOC75 and CAB were used for immuno blotting, as indicated (lower panels). C. YFP and chlorophyll fluorescence in fractions 1, 7, 11 and 21 was monitored by confocal microscopy. Scale bars: 1 μm (fractions 1–11) or 5 μm (fraction 21).