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Effect of Ltn1 mutations on nascent chain ubiquitylation (as in Fig. 7 B and D). Stalling reporter-encoding mRNA was translated in WT or Δltn1 extract, supplemented or not with recombinant Ltn1 (WT or mutants) (as indicated), and analyzed for ubiquitylation by denaturing IP. Lane 2 shows the “no Ltn1” control, and lane 8 shows the “no template” control reaction incubated with Ltn1 WT.
