Figure 3

Detection of myostatin propeptide by immunohistochemistry with antibody to the N-terminal. Mice were sacrificed at 12 months of age. Gastrocnemius (A to D) and biceps femoral (E to J) muscle samples were used for this study. Frozen muscle samples were cross-sectioned. Following the procedures of immuno-fluorescence double labeling, the sections were incubated with or without (negative control: A, C, E and H) primary antibody to myostatin propeptide. The P2 counter-stain mounting medium was applied, and the images were examined using Zeiss LSM5 laser-scanning confocal microscope. The nuclei were stained as red color, myostatin propeptide reactive with the primary antibody was showed as green color. A (wild-type) and C (transgenic): negative control (without primary antibody), showing myofibers and stained nuclei of Gastrocnemius. B: Gastrocnemius muscle section from wild-type mice was incubated with primary antibody, showing little myostatin propeptide on myofiber surface. D: Gastrocnemius muscle section from transgenic mice was incubated with primary antibody, showing intense myostatin propeptide on myofibers. E (wild-type) and H (transgenic): negative control (without primary antibody), showing myofiber and stained nuclei in cross-section of biceps femoral muscle. F and G: biceps femoral muscle cross-(F) and longitudinal (G) sections from wild-type mice, showing minimal reactive proteins to the primary antibody. I and J: biceps femoral muscle cross-(I) and longitudinal (J) sections from transgenic mice, showing strong and intense density of myostatin propeptide.