Fig. 3

The reporter mRNA with shortened Pou5f1 3′UTR is effectively translated in mouse mature oocytes. (A) A schematic diagram of the reporter assay in mouse oocytes. Immature oocytes were injected with reporter mRNAs and cultured in M2+ medium for 2 h, followed by incubation with M2+ or M2− for 14 hours. Immature (Im) and mature (M) oocytes were collected and extracted for the luciferase assay. (B) Schematic diagrams of reporter mRNAs with the full-length Pou5f1 3′UTR (Long) and 9- and 14-nt-deleted 3′UTRs (−9 and −14 nt). (C) Results of the luciferase assay (means ± SD; n ≥ 4). The activities relative to that of Long mRNA in immature oocytes. ***P < 0.001 and **P < 0.01, Dunnett’s test. (D) Schematic diagrams (top) and sequences (bottom) of reporter mRNAs carrying the full-length Pou5f1 3′UTR (Long) and 3′UTRs with 2-, 5-, or 9-nt mutations (2nt, 5nt, and 9nt mut, respectively). Sequences that are conserved in zebrafish and mouse are highlighted in gray. (E) Results of luciferase assay (means ± SD; n ≥ 3). The activities relative to that of Long mRNA in immature oocytes. ***P < 0.001, **P < 0.01, Dunnett’s test. n.s., not significant.