FIGURE 1.

Design of a morpholino antisense oligonucleotide (“α-SE”) that precludes the interaction between Su(H) and CUGBP1. (A) In vitro transcribed radioactive RNAs corresponding to the indicated regions of Su(H) 3′UTR (numbered from the stop codon) were incubated with no (lane 1) or increasing amounts (lanes 2–9) of recombinant CUGBP1. Free (F) and bound (B) RNAs were resolved by native electrophoresis and autoradiographed. (B) Sequence of Su(H) 3′UTR between nucleotides 495 and 631 (numbered from the stop codon). UGU trinucleotides are underlined, and the region complementary to α-SE morpholino is highlighted. (C) Radioactive RNAs corresponding to region 495–631 of Su(H) 3′UTR (upper panel) or Eg5 3′UTR (lower panel) were incubated with no (lane 1) or 50 nM (lanes 2–10) of recombinant CUGBP1, and increasing concentrations of α-SE morpholino (lanes 3–10). They were analyzed as in A. (D) CUGBP1 was immunoprecipitated from Xenopus egg extracts previously incubated with 5 μM of control (dark gray) or α-SE (light gray) morpholino. The indicated input and co-immunoprecipitated mRNAs were quantified by RT-qPCR. The percentages of mRNA recovered in the immunoprecipitated fractions are shown. Quantifications were in triplicate and error bars are the s.e. of the triplicates.