<i>In-silico</i> prediction of an ESE-enriched 25-bp sequence and identification of the interacting proteins.

Abstract

<p>(A) Schematic representation of the minigene construct containing the 75-bp <i>FGG</i> pseudoexon activated by the IVS6-320A>T mutation (indicated by an arrow). (bottom) ESE elements predicted by the ESEFinder program (<a href="http://rulai.cshl.edu/cgi-bin/tools/ESE3/esefinder.cgi?process=home" target="_blank">http://rulai.cshl.edu/cgi-bin/tools/ESE3/esefinder.cgi?process=home</a>) in the pseudoexon sequence are indicated by the histogram bars; the ESE-enriched 25-bp sub-region is underlined. (B) Western blot analysis of RNA-protein pulldown assays for the identification of <i>trans</i>-acting factors binding to the 25-bp region. (top) Sequences used in pulldown experiments. (bottom) Western blot analysis of hnRNP and SR proteins immunoprecipitated from HeLa nuclear extracts with either the wild-type or the scrambled 25-bp (as negative control) pseudoexon sequence.</p