Figure 1: Nested RT-PCR analysis of dystrophin expression in DMD patient cells treated with pseudo-exon-targeting 2OMe AOs (n=2); (a) The patient’s transcript amplicon containing the pseudoexon is 650 bp in length, and skipping of the pseudo-exon is generates a 578 bp product. Each AO (and the AO cocktail) was tested at a total concentration of 400 nM, 200 nM, 100 nM, 50 nM and 25 nM (shown left to right). UT: untreated patient samples. AO 1: Hi47A (-05+20); AO2: Hi47A (+22+46); AO 3: Hi47D (+11-14); -ve: no template RT-PCR control. (b) Densitometry was used to analyse the relative amounts of mutant and normal transcripts in treated and untreated cells. Additional overlapping AOs were designed to anneal to the cryptic acceptor splice site, as shown in Figure 2.