Fig. 3: Puf6 is enriched in the nucleolus and associates with early 60S pre-ribosomes.

a Yeast cells expressing Puf6-GFP and the nucleolar marker Gar1-mCherry were grown to mid-log phase in YPD and then analyzed by fluorescence microscopy. Scale bar = 5 µm. b Cells expressing Puf6-GFP, Arx1-GFP, or Gar1–GFP were mated with a kar1-1 strain expressing Nup82-mCherry. The resulting heterokaryons were analyzed by fluorescence microscopy. Arx1-GFP and Gar1–GFP serves as a positive and negative control for the shuttling assay, respectively. Scale bar = 5 µm. c TAP-eluates of the early to late 60S pre-ribosomes isolated via the indicated TAP-baits were separated on a NuPAGE 4–12% Bis-Tris gradient gel and subjected to Silver staining or Western blotting using antibodies directed against Puf6, Nog2, and Nmd3. The r-protein uL29 (yeast Rpl35) served as a loading control. WT-BY4741 (untagged strain) served as the negative control. d The Puf6-TAP particle was separated on a NuPAGE 4–12% Bis-Tris gradient gel and analyzed by Silver staining. The indicated proteins were identified by mass spectrometry. e Puf6-TAP and Nog2-TAP eluates were separated on a NuPAGE 4–12% Bis-Tris gradient gel and subjected to Western blotting using the indicated antibodies. The r-protein uL29 (yeast Rpl35) served as a loading control. Source data are provided as a Source Data file.