Fig. 2

Immunoblot investigation of R16/17-Syn PDZ fusion protein expression and restoration of sarcolemmal nNOSμ in mdx mice. a. Whole muscle lysate western blot. The robust expression of the fusion protein was revealed by antibodies against R17, GFP, and the syntrophin PDZ domain. The total amount of nNOS in the AAV-injected muscle is almost the same as that of uninjected mdx muscle. Quantification of band intensity confirmed that the total amount of nNOS in the wild-type muscle is significantly higher than in mdx and AAV-injected muscles (asterisk, significantly different from mdx or AAV-injected muscle). GAPDH is the loading control. b. Microsomal preparation western blot. The membrane expression of the fusion protein was detected by antibodies against R17, GFP, and the syntrophin PDZ domain. Restoration of sarcolemmal nNOSμ by the fusion protein was confirmed by identifying nNOS in the microsomal preparation of AAV-injected muscle. ATPase is the loading control. Open arrowhead, full-length dystrophin; filled arrowhead, R16/17-Syn PDZ fusion protein; arrow, endogenous syntrophin