Fig. 3

Simvastatin treatment does not attenuate inflammation and fibrosis in gastrocnemius of mdx mice. A Representative pictures of hematoxylin and eosin (H&E) staining of the gastrocnemius muscle with semi-quantitative analysis of inflammation; scale bar: 100 μm; mean ± SEM; n=5–6/group. B Unaffected by simvastatin treatment expression of Hmox1 gene in gastrocnemius, presented as a mean ± SEM; n=10–13; qRT-PCR. C Necrosis assessment using immunofluorescent staining of IgM/IgG/IgA binding (green) with laminin (red) and its calculation presented as a mean ± SEM; n=6–13/group; scale bar: 100 μm. D Representative photos of Masson’s trichrome staining with semi-quantitative analysis of collagen deposition showing no changes in the extent of fibrosis in gastrocnemius of simvastatin-treated animals; scale bar: 100 μm; n=5/group. E Unaffected by simvastatin treatment expression of Spp1 gene in gastrocnemius, presented as a mean ± SEM; n=10–14; qRT-PCR. F The protein level of serum marker of fibrosis, OPN, n=10–13/group, mean ± SEM; ELISA. G Unaltered by the treatment expression of fibrotic markers: Tgfb1 and Mmp11, in gastrocnemius of mdx mice and a significant decrease in Col1a1 mRNA; n=12–14/group, WT level marked with the dotted line; qRT-PCR. Data are presented as mean ± SEM; ^*for mdx vehicle vs. WT and # for mdx simvastatin vs. mdx vehicle comparison; ^*p < 0.05, ^**p < 0.01, ^***p < 0.001, ^****p < 0.0001, and ^#p < 0.05