Fig. 3

Induction of F-α-DG in cardiac and skeletal muscles of P448L mice treated with ribitol for 6 months. Seven-week-old P448L mice were given drinking water only (n = 4), or drinking water supplemented with 5% ribitol (n = 4). a IIH6C4 immunohistochemical staining of cardiac (heart), tibialis anterior (TA), and diaphragm tissues from either untreated or 5% ribitol-treated P448L mice (left and middle panel, respectively), and C57 mice. Nuclei were counterstained with DAPI (blue). Arrows indicate the revertant fibers expressing F-α-DG. Arrowhead indicates the degenerating fibers and focal accumulation of nuclei. Scale bar, 50 μm. b Western blot and laminin overlay assay of lysates from heart, TA, and diaphragm (diaph) of two untreated (−) or two ribitol-treated (+) P448L, and C57 mice. F-α-DG was detected by blotting with IIH6C4 and by laminin overlay assay (Laminin OL). Core of α-DG was detected by AF6868 antibody with weaker signals for the ribitol-treated samples. Detection of α-actin was used as a loading control. Arrowheads indicate laminin-binding bands. The upper band with laminin-binding assay are endogenous laminin present in all samples. c Quantification of IIH6C4 band intensity from western blot. Values were normalized to α-actin expression for each tissue and presented as percentage of C57 levels. Error bars represent mean ± SEM. Unpaired t test. *p ≤ 0.05