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. Author manuscript; available in PMC: 2020 Aug 3.
Published in final edited form as: Sci Transl Med. 2019 Nov 27;11(520):eaat6072. doi: 10.1126/scitranslmed.aat6072

Fig. 2. Evaluating calpain 3 gene transfer in the Dysf−/−Capn3−/− dKO model.

Fig. 2.

(A) The expression cassette used in this study includes a human desmin promoter, a chimeric β-globin intron, the mfa calpain 3 coding sequence fused with a V5 tag, and a simian SV40 polyadenylation signal. In addition, a tandem miR-208a target sequence was included in the 3′UTR of the vector (rAAV-C3+miRT). (B) Schematic representing design of the in vivo experiment (n = 4 to 7). IM, intramuscular; D, days. (C) Calpain 3 mRNA expression obtained by ddPCR in skeletal muscle extracts. Statistical analyses were performed using nonparametric Kruskal-Wallis test and the post hoc multiple comparison Dunn’s test (n = 4 to 7). **P < 0.01. (D) Representative gluteus muscle sections stained by hematoxylin phloxine saffron (HPS), Sirius red (SR), myosin heavy chain developmental form (MHCd), and CD11b. Scale bars, 150 μm. (E) Quantitative RT-PCR (qRT-PCR) quantification of genes representative of several dystrophic features: Cd11b for macrophage infiltrates, myogenin for regeneration, and collagen VI (Col6a) for fibrosis in muscles of Blaj, BDC HOHO, and injected BDC HOHO. The data are expressed as abundance normalized by Rplp0. Statistical analyses were performed using nonparametric Kruskal-Wallis test and the post hoc multiple comparisons Dunn’s test (n = 4 to 7) *P < 0.05; **P < 0.01. (F) Radar graph showing recovery score for each dystrophic markers. Values are expressed relative to those of BlaJ mice (set to 100%) and BDC HOHO (0%). Individual data are presented in fig. S1B.