Fig. 6

Myogenic reprogramming of DMD patient-derived urine cells. a Schematic showing organization of dystrophin exons 42 to 51 (top) and DMD patient with a frameshift deletion of exons 46 and 47 (bottom). RT-PCR demonstrated DMD transcript expression from iMyoD-transduced line 11 control and DMD ex46/47del mutant cell lines, after tamoxifen induction (2.5 μM, 24 h) and differentiation as indicated. Blue arrows indicate the location of the outer primers that produce the PCR products in the gel below. Red arrows indicate nested primers used to generate transcripts used for sequencing. b Sequencing results from RT-PCR products from iMyoD-reprogrammed urine-derived cells. The sequenced transcripts were generated from a set of nested primers (red) shown in a. Line 11 showed the expected exon45-exon46 and exon47-exon48 junctions. The ex46/47del DMD individual shows the out-of-frame mutant exon45-exon48 junction. c Immunofluorescence images of iMyoD line 17 and DMD ex46/47del cells labeled with dystrophin using the NCL-DYSB antibody to dystrophin. The antibody to dystrophin showed membrane localization of dystrophin in control cells (top). No dystrophin was detected in DMD patient-derived myotubes. Scale bar = 50 μm