Restoration of dystrophin protein expression by AO88.
(a) Transfection procedure for antisense oligonucleotide (AO) 88 in induced skeletal muscle cells. DMD-Myocytes were transfected with AO88 at day 7. Protein and RNA extraction was performed after 48 hours of transfection. (b) Exon 45 skipping was assessed by RT-PCR to compare Control-Myocytes (Father and B7) to DMD-Myocytes (Δ44 and Δ46–47). Exon 45 skipping led to an in-frame shift of exon 46 with Δ44 and exon 48 with Δ46–47, which resulted in smaller bands. (c) Western blotting analysis detecting the C-terminus of the dystrophin protein. AO-treated DMD-Myocytes showed restored expression of dystrophin protein. (d) Merged immunocytochemistry images for dystrophin and DAPI. Restored dystrophin protein expression was detected when samples were treated with AO88 (+AO). Scale bar, 20 μm.