Restoration of dystrophin and its associated proteins at the sarcolemma of mdx muscles by genome-editing. (a) Western blotting of gastrocnemius muscle homogenates from wild-type (WT), mdx, and mdx with adenoviral vectors carrying GFP-2A-cas9 and gRNA using anti-dystrophin and anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibodies. 22, 11, and 5.5 µg total proteins from the WT muscles were loaded per lane. (b) Quantitative analysis of western blotting in WT, mdx, and mdx injected with adenoviral vectors. (c,d) Confocal immunofluorescence images of dystrophin (c, red) and neuronal nitric oxide synthase (nNOS) (d, red) in muscle cryosections treated with or without EGFP-2A-cas9/gRNA adenovirus. The images are representative of five experiments. Scale bar: 100 µm. ***P < 0.001.