Skip to main content
. 2008 Jun 16;181(6):935–944. doi: 10.1083/jcb.200801181

Figure 4.

Figure 4.

Nsa1 is associated with mature 60S ribosomes in the rix7Δ14N mutant. (A) Affinity purification of Nsa1-TAP from wild-type RIX7 or rix7Δ14N mutant cells that were first grown at 23°C and then shifted for 3 h to 30°C. The final EGTA eluates were analyzed by SDS-PAGE and Coomassie staining (top) and Western blotting using anti-Nop7, anti-Nog1, anti-CBP, anti-Rpl3, anti-Rpp0, anti-Rpl10, and anti-Rps8 antibodies (bottom). The band marked with an asterisk corresponds to Rpp0. The bands corresponding to ribosomal proteins are indicated. (B) rRNA composition of Nsa1-TAP affinity-purified from wild-type RIX7 or rix7Δ14N mutant cells that were first grown at 23°C and then shifted for 3 h to 30°C. Aliquots of total RNA and of the final EGTA eluates were resolved on a 1.5% agarose-formaldehyde gel, transferred to a nylon membrane, and analyzed by Northern blotting using the indicated probes to detect pre-rRNA and mature rRNA species. (C) Affinity purification of mature 60S ribosomal subunits via Rpl24A-TAP from wild-type RIX7 or rix7Δ14N mutant cells expressing Nsa1-GFP. Cells were first grown at 23°C and then shifted for 3 h to 30°C. The final EGTA eluates were analyzed by SDS-PAGE and Coomassie staining (top) and Western blotting using anti-GFP, anti-CBP, anti-Rpl3, anti-Rpp0, and anti-Rps8 antibodies (bottom). The bands marked with asterisks contain Stm1, Rpl5, Rpp0/Asc1, and Rpl2/Rps1/Rps3/Rps4/Rps0/Rpl8/Rps6, respectively. MW, molecular weight protein standard.