Figure 2.

In vitro proof of principle of antisense oligonucleotides (AON)-mediated ALK5 exon skipping. (a) AONs targeting exon 2 of ALK5 (ALK5 AON) were transfected at different concentrations into C2C12 mouse myoblasts. Two days after transfection, the efficacy to induce exon skipping was assessed by reverse transcription PCR ALK5-specific primers in flanking exons (red arrowheads). Nontransfected (NT) cells or cells transfected with control AON (Ctrl AON, nontargeting) served as controls. (b) Sequencing of excised PCR products showed exclusion of the targeted exons ALK5 AON transfection (exon 2 skip) in C2C12 cells. (c) Quantitative real-time PCR of C2C12 samples was performed to compare full-length ALK5 transcript expression using primers in the skipped exon and in the flanking exon. Cells were transfected with 200 nmol/l AON. These data are shown as the average of at least three independent experiments and is shown relative to control AON samples. Error bars represent standard deviations. *P < 0.05; **P < 0.01