Fig. 5
From: Direct reprogramming of urine-derived cells with inducible MyoD for modeling human muscle disease
Myogenic reprogramming of LGMD2C UDCs. a Schematic showing exon organization of the γ-sarcoglycan (SGCG) mRNA transcript and two LGMD2C patients with frameshifting deletions affecting the SGCG locus that lead to premature stop codons (red triangles). One has a deletion of exons 5 and 6 (ex5/6del), and the other has a deletion of exon 7 (ex7del). RT-PCR demonstrated SGCG transcripts from UDCs after treatment with tamoxifen (2.5 μM, 24 h) and culture in differentiation media for 28d. Orange arrowheads indicate the mutant transcripts lacking the expected exons. b Real-time semi-quantitative PCR results demonstrate myogenic reprogramming of ex5/6del (top) and ex7del (bottom) after treatment with tamoxifen (2.5 μM, 24 h) and culture in differentiation media as indicated (7, 14, and 28 days). c Images of myogenic markers in iMyoD control and LGMD2C ex5/6del and ex7del patient-derived lines. LGMD2C patient-derived cells formed myotubes comparable to those of control cells. Nuclei (blue); Scale bar = 50 μm. d Desmin, α-actinin, dystrophin, and fast MHC were expressed in LGMD2C cells. γ-sarcoglycan (green) was not detected. White boxes outline double-labeled regions of interest (ROI) enlarged in far right panels. Images show γ-sarcoglycan expression and localization to the membrane in line 11, as opposed to no detectable expression of γ-sarcoglycan in LGMD2C patient myotubes. Scale bar = 100 μm in left and middle panels; 50 μm in right panels