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. 2014 Mar 17;204(6):909–917. doi: 10.1083/jcb.201308139

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© 2014 Ossareh-Nazari et al.

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Figure 2. — The 60S ribosomal protein Rpl25 is a substrate of both Ltn1 and Ubp3. (A) GFP or Rpl25-GFP were expressed in cells transformed (+) or not transformed (−) with a plasmid encoding 6His-ubiquitin. (top) Expression levels of GFP and Rpl25-GFP in whole cell lysates were verified with anti-GFP blotting. (middle) Purified 6His-ubiquitin conjugates were examined by anti-GFP blotting. (bottom) 6His-ubiquitin expression and efficiency of purification was controlled using an anti-6His antibody. White lines indicate that intervening lanes have been spliced out. (B) Analysis of ubiquitylated forms of Rpl25-GFP in wild-type and the indicated mutant cells as in A. (C) Expression of Rpl25-GFP and its modified form were analyzed by anti-GFP blot of extracts from wild-type and mutant cells starved for the indicated periods. Ub, ubiquitin; WT, wild type.