Fig. 7
Miniagrin treatment restores expression of dystroglycan receptor in muscles. a Cryosections of the tibialis anterior muscle from dy2J mice untreated (saline) or treated with MABs + mMAG double stained with anti-myc (recognizing mMAG) and anti β-dystroglycan integrin antibody. The immunofluorescence shows that myc staining is present only in treated muscle. Staining for β-dystroglycan is more intense and diffuse in treated muscle. Images are acquired by confocal microscope with the same z-stack and laser intensity. DAPI staining identifies nuclei. b Western blot analysis of tibialis anterior muscle from dy2J mice untreated, or treated with MABs+mMAG or MABs alone, and and wild type (Wt) controls, stained with anti β-dystroglycan, and β-tubulin as loading control. Quantification of western blot analysis is reported as an average of 3 independent experiments and represented as the ratios β1 integrin/actin, assigning Wt as 1. A significant decrease of β-dystroglycan (n = 3) is present in muscle of dy2J mice as compared to Wt control; the amount of β-dystroglycan is significantly increased after treatment with MABs+mMAG (n = 3). c Cryosections of wild type (Wt), dy2J untreated (saline), dy2J treated with MABs, and dy2J treated with MABs + mMAG stained with anti α-dystroglycan antibody and DAPI. Staining for α-dystroglycan was increased in dy2J mice as compared to Wt and more similar to Wt control in dy2J mice treated with MABs or MABs + mMAG. Western blot analysis confirmed the significant increase of α-dystroglycan in dy2J, as compared to Wt mice; dy2J mice treated with MABs or MABs + mMAG did not show further changes. d Western blot analysis of tibialis anterior muscle from dy2J mice untreated, or treated with MABs+mMAG or MABs alone, and Wt controls, stained with anti α-dystroglycan, and β-tubulin as loading control. Quantification of western blot analysis is reported as an average of 3 independent experiments and represented as the ratios β1 integrin/actin, assigning Wt as 1. A significant increase of α-dystroglycan (n = 3) is present in muscle of dy2J mice as compared to Wt control; similar levels are also observed in dy2J mice treated with MABs (either alone or with mMAG; n = 3). Scale bar = 50 μm in (a) and 100 μm in (c). Student t test; *p < 0.05; **p < 0.01; error bars indicate SEM