hnRNP A1 and hnRNP F Modulate the Alternative Splicing of Exon 11 of the Insulin Receptor Gene
Figure 5
hnRNP A1 but not hnRNP F binds at the junction of exon 11 and intron 11 to repress exon inclusion.
Panel A: The nucleotide sequence of the RNA templates of the wild type exon 11/intron 11 junction and the -3U→C mutation (underlined). Binding sites for SRSF1 and hnRNP A1 are indicated by bars above. Representative blots show the results of in vitro RNA pull-down assays followed by immunoblotting for hnRNP A1, F, H, and U1-70K and SRSF1. Panel B: The nucleotide sequence of the RNA templates of the wild type exon 11 and the LS5 mutation (underlined). Representative blots show the results of in vitro RNA pull-down assays followed by immunoblotting for hnRNP A1 and SRSF1. Panel C: Transient transfection of the wild type minigene hIR and the -3U→C mutant in the absence and presence of the expression vector of hnRNP A1 or CELF1. A representative gel is shown. The ratio of isoform B over isoform A of n = 3 experiments with ± SD is shown. Asterisks indicate statistical significance vs. wild type (** p≤ 0.01).