As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re-organized for online delivery, but are not copy-edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.

emmm201302621.reviewer_comments.pdfPDF document138KReview Process File
emmm201302621-sm-0002-SuppFig-S1.pdfPDF document505K

Figure S1. Further in vivo efficacy data from the pre-symptomatic mdx trial. (A) The drop in force for EDL muscle after 10 lengthening contractions ex vivo showed a significant reduction upon VBP15 treatment at 15 mg/kg. (B and C) Live-animal imaging of cathepsin protease activity via ProSense680 shows reduced inflammatory disease in the forelimbs of VBP15-treated mdx mice (B) images; C quantitation of fluorescence; n ≥ 6 mice/group). (D) H&E stained histology of diaphragm muscle shows a decrease in calcified myofibres upon VBP15 treatment. In diaphragm sections, calcified fibres were present in a majority (5 of 6) of vehicle mdx mice. VBP15 treated mice showed reduced calcification upon quantitation, with only 1–2 mice per group exhibiting any calcified fibres (n = 6 mice/group). (*p < 0.05, n = 10 mice/group).

Figure S2. Further in vivo immunology and side effect data from the pre-symptomatic mdx trial. (A) Spleen to body mass ratios were measured upon trial conclusion (8 weeks). Note, mdx spleens were enlarged in comparison to WT, while VBP15 normalized spleen size toward WT levels and prednisolone reduced spleen size such that it was lower than WT. (B) Peripheral blood leucocyte counts were assayed at trial conclusion. (C–E) Histological analysis of gastrocnemii muscle shows increased skeletal muscle degeneration and fibrosis in prednisolone-treated mice, but not VBP15 mice. A significant increase in degenerating fibres (C) was present in prednisolone treated gastrocnemii, quantified in H&E stained sections. Digital quantitation (D) of sirius red staining shows a significant increase in skeletal muscle fibrosis in prednisolone treated mice. Representative images of sirius red staining are provided (E), to the right of the panel is a higher magnification image from the area outlined in box. (n ≥ 12 mice/group for (A and B); n = 6 mice/group for (C and D); *p < 0.05, **p < 0.05, ***p < 0.05).

Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.