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Fig. 1 Establishment of a ribosome affinity purification system in zebrafish. (A) Scheme of a FLAG-tagged rpl36 gene locus. Genome sequences around the stop codon of wild-type (WT) and rpl36-FLAG strains are shown. The sequence chromatogram of rpl36-FLAG embryos is indicated below. (B) Representative polysome profiles of rpl36-FLAG embryos at 24 hpf. The distribution patterns of ribosomal proteins are indicated below. (C–E) Validation of the ribosome purification system with WT and rpl36-FLAG embryos at 24 hpf. Total lysates (input) and FLAG-immunoprecipitants (IPs) were subjected to RNA electrophoresis (C), immunoblotting analysis using antibodies against the indicated proteins (D), and protein staining (E). (F–H) Validation of the ribosome purification system during various developmental stages. The developmental time points are indicated above as hpf. Total lysates (input) and FLAG-IPs were subjected to RNA electrophoresis (F), immunoblotting analysis using antibodies against the indicated proteins (G), and protein staining (H).