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. Author manuscript; available in PMC: 2009 Dec 12.
Published in final edited form as: Biochem Biophys Res Commun. 2008 Oct 16;377(2):653–657. doi: 10.1016/j.bbrc.2008.10.029

Figure 1. Endogenous ElrA and AUF1 bind cyclin B2 3′UTR.

Figure 1

A, UV-crosslink analysis of 4 and 8h embryos with radiolabeled cyclin B2 3′UTR in the presence of 0, 1, 5, or 10 molar excess unlabeled cyclin B2 3′UTR or cyclin A1ΔARE. Arrow, 37-kDa protein competed by B2 3′UTR. Arrowhead, proteins not competed. Double arrowheads, proteins competed by both. B2, cyclin B2 3′UTR; B2R, cyclin B2 3′UTR+RNase. B, Immunoblot of unfertilized eggs (U) through 10 hpf embryos sequentially probed with ElrA and AUF1 antibodies. Top numbers, hours postfertilization. C, UV-crosslink of 4 and 8h embryos with radiolabeled cyclin B2 3′UTR or A1ΔARE (A1) followed by immunoprecipitation with IgG, αElrA or αAUF1. Input, UV-crosslink before immunoprecipitation. Arrowheads, immunoprecipitated protein. D, RNA-pulldown assay of 4 and 8h embryos with biotinylated cyclin B2 3′UTR; followed by immunoblotting for AUF1, ElrA, and CPEB (Pulldown). Input, immunoblot of embryos used for the pulldown. 8+a, embryos injected with α-amanitin. E, RNA-pulldown of 4 and 8h embryos with biotinylated cyclin B2 3′UTR or A1ΔARE; followed by immunoblotting for AUF1, ElrA, and actin. Markers are in kDa for A-C; n=3 for A, C-E; n=5 for B.