Figure 3.

Effect of AMOs against Region 147–286 on Exon 2 Inclusion in the MUT Minigene Context

(A) Schematic representation of AMOs 1, 2, and 3 targeting the 147–286 region of GAA exon 2. The AMOs’ target sequences are indicated by black lines. (B) RT-PCR analysis of HeLa cells co-transfected with AMOs 1, 2, and 3 and the GAA MUT minigene. AMOs were delivered as a single unit or in different combinations to a final concentration of 15 μM. Cells transfected with the mutated minigene, together with Endo-Porter reagent, were used as internal control; not transfected (NT) cells were used as RT-PCR control. The agarose gel picture shows a representative result of three independent experiments. (C) qRT-PCR analysis of GAA exon 2 inclusion (N) in cells co-transfected with AMOs (1, 2, and 3) and the GAA MUT minigene alone. The absolute abundance of the N form is normalized on AlfaGlobin expressed as the fold of increase over the minigene alone. qRT-PCR data are represented as mean ± SEM of three independent experiments in a histogram plot (*p < 0.05, ***p < 0.001).