Figure 4.

Further Analysis of 12-Week Gene Therapy Experiment in BlAJ/NRG Mice

(A) Immunohistochemistry staining on G, H, and Q muscle sections harvested 12 weeks after vascular injection of HBSS alone, pDYSF, or pDYSF-FST. Laminin is stained red, and dysferlin is stained green. A section from the positive control NRG mouse is also shown. Bar, 50 μm. (B) Presence of plasmid DNA was detected by PCR in muscles injected with HBSS alone, pDYSF, or pDYSF-FST. Left diagram shows the position of the primers used to detect an identical 1,242-bp region of DNA present on pDYSF and pDYSF-FST. Representative PCR lanes show DNA extracted from Q, H, and G muscles from legs treated with either HBSS alone or pDYSF-FST. Control lanes are B = BlAJ/NRG, NRG; B+D = BlAJ/NRG + pDYSF; and B+F = BlAJ/NRG + pDYSF-FST. (C) Follistatin concentration was quantified by ELISA from muscles injected with HBSS, pDYSF, or pDYSF-FST. Legs injected with pDYSF-FST expressed significantly more follistatin than legs injected with HBSS alone or pDYSF (pooled for statistical purposes) 28 days after the injections. (D) Concentration of Evan’s blue dye was significantly lower in hamstring muscles of legs injected with pDYSF-FST compared to legs injected with HBSS alone or pDYSF 12 weeks after injections. Variability was also significantly lower in muscles injected with pDYSF-FST compared to HBSS alone or pDYSF-injected muscles, as determined by F test. Q, quadriceps; H, hamstring; G, gastrocnemius. Data are mean ± SEM, with n = 4–8 and *p < 0.05.