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Mutations that stabilize the stem-loop structure reduce U2AF65 binding affinity and repress exon 5 inclusion in vivo. (A) Schematic of different mutations, the thermal stability of each mutation and the binding affinity of U2AF65 for each RNA. (B) Binding curve of tested mutations. (C) Representative RT-PCR data from in vivo splicing of cTNT mutant series. (D) Quantitation of RT-PCR data, and data from CUG960 cotransfection experiments.
