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. Author manuscript; available in PMC: 2013 Mar 20.
Published in final edited form as: Neuron. 2012 Sep 20;75(6):1067–1080. doi: 10.1016/j.neuron.2012.07.009

Figure 4. In vitro selection of nElavl binding RNA molecules.

Figure 4

(A) Representative results from in vitro RNA selection with the nElavl proteins. RNA selection was carried out using the Elavl2, Elavl3 or Elavl4 proteins for 6-8 rounds of selection (R6-R8, as indicated). Consensus GU rich elements are shown in blue and below the diagram. (B) RNA gel-shift assay, in which DR8-9 clone RNA selected by Elavl4 was incubated with recombinant Elavl4 as indicated. Multiple forms of Elavl4/RNA complexes (arrows) have slower migration profiles with increasing amounts of protein (RNA=50 fmol/lane, Protein=0, 25, 50 or 75 ng), and this effect is specific, as no effect on RNA mobility was seen with either hnRNP A1 (25 ng), or when Elavl4 was incubated with an irrelevant control RNA (SB2, a 52 base NOVA1 consensus sequence). (C) Results of filter binding assays in which the indicated amounts of Elavl4 fusion protein were incubated with radiolabeled selected RNA (where the number of GUUGU repeats (n) is shown, see (A)). Red: DR8-2 RNA; Blue: DR8-15 RNA; Green: DR7-5 RNA; Black: control SB2 RNA (50 fmol/reaction). Estimated kD’s are shown.