Fig. 2: Cdc48-Ufd1-Npl4 is recruited to stalled ribosomes by Ltn1’s elbow ___domain.

a Positioning of Cdc48-Ufd1-Npl4 relative to the 60S ribosome. Focused view on bottom shows a 2D class average of sub-extracted Cdc48 complexes from RQC particles. b Close up view of the Ltn1-Cdc48 interface at the Ltn1 elbow. Key interaction residues are labeled. Residues mutated to generate the Ltn1^AAA mutant are indicated with a red outline. c Silver stained SDS-PAGE of RQC co-IPs expressing WT or Ltn1^AAA. d Anti-Cdc48 immunoblot from RQC co-IPs. Anti-FLAG (Rqc1) blot shown as loading control. e Immunoblot of whole cell lysates expressing nonstop substrate (Protein A-nonstop) in Ltn1∆ cells and re-expressed with either wild-type Ltn1 or Ltn1^AAA mutant. Anti-Pgk1 blot shown as loading control. In (c–e), each experiment was repeated at least three times as independent biological replicates.