Fig. 3.

Csy4*-based RNA affinity purification isolates known RNA-binding proteins. (A) Comparison of Csy4*-based and biotinylated RNA purification for pre-let-7a. Both transcripts were tagged at the 5′ end. Eluted samples were separated via 4–20% SDS/PAGE and visualized by silver staining. (B) Csy4 hairpin RNA-tagged pre-let-7a copurifies with Lin28A. Csy4 hairpin RNA-tagged pre-let-7a and pre-miR-21 were used as bait for Csy4*-based RNA affinity purification from HeLa and NT2 cellular extracts. Pull-down eluents were analyzed by SDS/PAGE and visualized by silver staining (Left). (C) Western blots of Csy*-based pull-down eluents from B using an anti-Lin28A antibody.