Skip to main content
. 1999 Feb 8;144(3):389–401. doi: 10.1083/jcb.144.3.389

Figure 9.

Figure 9

L25-GFP containing ribosomes isolated from nup49-313/L25-GFP and Gsp1p-G21V/L25-GFP cells. Ribosomes were isolated under low salt conditions (100 mM KCl) from (A) nup49-313/L25-GFP cells shifted for 14 h to 33°C and incubated for another 4 h at 20°C and (B) L25-GFP cells expressing Gsp1p-G21V under restrictive conditions (i.e., galactose medium) as described in the legend of Fig. 5 and in Materials and Methods. Whole cell extracts were prepared, loaded on a 10–40% sucrose gradient and centrifuged for 12 h at 150,000 g. The fractions from the sucrose density gradient were TCA-precipitated, resuspended in SDS-sample buffer and analyzed by SDS-PAGE and Coomassie-staining (upper part) or Western blotting using anti-GFP antibodies (lower part). The position of L25-GFP is indicated. The asterisks mark the position of prominent ribosomal proteins. Marker proteins (10-kD ladder with a stronger 50-kD band) are also depicted.