Figure 1: Expression of dystrophin and myogenic markers in GSΔ44 biopsy and primary myoblasts.

(a,b) Western blot on proteins (20 μg) extracted from human muscle biopsies of a healthy control (CTRL) and from GSΔ44 probed with antibodies against dystrophin (DYS, a), or myogenin (MYOG), MHC and muscle creatine-kinase (MCK, b). Actinin (ACTN) was used as a loading control. (c) Quantitative RT–PCR of myogenic markers and myomiRs performed on RNA extracted from healthy control (CTRL) and GSΔ44 muscle biopsies. Bar plots show relative expression levels with respect to control set to 1: values were normalized with GAPDH and hypoxanthine phosphoribosyltransferase (HPRT) for mRNAs and with U6 and miR-16 for miRNAs. Bars indicate a confidence level of 95%. (d) Western blot on proteins extracted from healthy control (CTRL) and GSΔ44 primary myoblasts upon 10 days of shift to differentiation conditions and probed with antibodies against dystrophin (DYS), myogenin (MYOG) and MHC. The amount of proteins loaded is indicated below each lane. Actinin (ACTN) was used as a loading control. (e) Nested RT–PCR performed on RNA from biopsy samples (upper panel) and differentiated myocytes obtained as in d (lower panel); 20 ng of cDNA were amplified with primers located in exons 42 and 53 and then with primers in exons 43 and 46 (upper panel) and in exons 43 and 48 (lower panel). A schematic representation of the WT, interrupted and restored ORFs is shown on the side. M, molecular weight marker: GeneRuler 100 bp (Thermo Scientific) for the upper panel and GeneRuler 1 kb (Thermo Scientific) for the lower panel; sizes are shown on the side.