FIG. 1.

S-nitrosylation extent and skeletal muscle mass in GSNOR-KO mice. (A) Evaluation of protein S-nitrosothiols (PSNOs) amount in total homogenates of tibialis anterior of 2-month-old GSNOR-KO (KO) and wild-type (WT) mice, subjected to biotin-switch assay and revealed by incubation with horseradish peroxidase (HRP)-conjugated streptavidin. Lactate dehydrogenase (LDH) was selected as a loading control from a set of elective “housekeeping” proteins on the basis of the coherence between immune-reactive band signal and nitrocellulose Ponceau-Red staining. (B) Western blot analyses of neuronal nitric oxide synthase (nNOS), inducible NOS (iNOS), and endothelial NOS (eNOS) performed in tibialis anterior homogenates. LDH was selected as a loading control. (C) Representative fluorescence microscopy images of tibialis anterior sections from KO, WT, and dystrophin-null (mdx) mice on staining with anti-nNOS (green), and DAPI (blue) to highlight nuclei. Scale bar, 40 μm. (D) Representative H&E stained sections of gastrocnemius of KO and wild-type WT mice. Centro-nucleated myofibers areas including small-sized fibers are indicated (arrowheads and white dotted lines, respectively). Scale bar, 100 μm. (E) Representative picture displaying paws detail of KO and WT mice. Dotted areas, muscle volume of WT legs. (F) Weight measures of tibialis anterior and gastrocnemius on tendon-to-tendon isolation from KO and WT mice. Results shown are the means±SEM of n=6 animals for each group, and the indicated p-value was calculated with regard to WT. (G) Overall body weight of KO and WT mice. Results shown are the means±SEM of n=6 animals for each group, and the indicated p-value was calculated with regard to WT. To see this illustration in color, the reader is referred to the web version of this article at www.liebertpub.com/ars