Fig. 1.

U2AF⁶⁵ stimulates exon 7 exclusion in both SMN2 and SMN1 pre-mRNA. (A) RT-PCR results of both endogenous SMN1 and SMN2 pre-mRNA are shown from untreated, nonsilencing shRNA-treated and U2AF⁶⁵-shRNA-treated 293A, C33A, SH-SY5Y, and GM03813 cells. The percentage of exon 7 included RNA versus total RNA and its SD are indicated at the bottom. Immunoblotting analysis of these cells using an anti-U2AF⁶⁵ (MC3) antibody are shown. (B, Left) The scheme of SMN2 minigene is shown with the intron RNA as a thicker line, whereas the vector sequence as a dot line. The RNA sequence of pseudo 3′ splice-site is shown. (Right) Shown is RT-PCR analysis from cells that express SMN2 minigene with overexpression of Flag-U2AF⁶⁵ plasmid or control plasmid. Immunoblotting analysis with antiflag antibody is illustrated. (C, Left) SMN1 minigene scheme is shown. (Right) Results of RT-PCR analysis of the SMN1 minigene.