Journal: Science Advances

Article Title: Lysosomal damage is a therapeutic target in Duchenne muscular dystrophy

doi: 10.1126/sciadv.adv6805

Figure Lengend Snippet: ( A ) Schematic representation of the study setup. Six-week-old Dmd mdx mice were injected intravenously with a rAAV encapsulating a μDys encoding sequence, under the control of an spc512 promoter, at two doses of 5 × 10 12 vg/kg (low dose) and 1 × 10 13 vg/kg (high dose). Four-week-old Sgcg −/− mice were injected intravenously with a rAAV9 encapsulating the human sequence of SGCG, under the control of a desmin promoter, at a dose of 2 × 10 13 vg/kg. IV, intravenous. ( B ) Dosage of CK in serum of mice at the end of the study (average ± SD). ( C ) ELISA quantification of MYOM3 in the serum of the mice before euthanasia. Data are presented as the scatterplot (average ± SD). ( D ) Global force evaluation by an escape test (force in newton normalized to body weight in grams) before euthanasia of mice (average ± SD). ( E ) Relative Lgals3 mRNA expression in the GA normalized to Rplp0 (average ± SD). ( F ) Representative confocal images of transversal sections of GA muscle immunostained for LAMP2 and LGALS3. Scale bars, 25 μm. ( G ) Quantification of double-positive puncta (LAMP2 + LGALS3 + ) in GA myofibers (median and range shown). An ANOVA test with Tukey’s multiple comparisons test was used for statistical comparisons. * P < 0.05, ** P < 0.05, *** P < 0.001, and **** P < 0.0001. Gray asterisks represent relative comparison to the “WT_Phy Ser” group, and red asterisks represent comparison to the “mdx_Phy Ser” group.

Article Snippet: An anti-MYOM3 polyclonal capture antibody (Proteintech Lab, ref. 17692-1-AP) was coated to the wells of a multiarray plate containing electrodes (MSD, ref. L15XA-3) (overnight, 4°C).

Techniques: Injection, Sequencing, Control, Enzyme-linked Immunosorbent Assay, Expressing, Comparison

Journal: Science Advances

Article Title: Lysosomal damage is a therapeutic target in Duchenne muscular dystrophy

doi: 10.1126/sciadv.adv6805

Figure Lengend Snippet: ( A ) Study setup. Dmd mdx-4Cv mice were started on TR treatment (2% dilution in drinking water) at 3 weeks. For groups 4 and 5, mice were injected intravenously with a rAAV9 encapsulating the μDys sequence at a dose of 7 × 10 12 vg/kg. ( B ) Seric CK analysis before the escape test (average ± SD, n = 5). ( C ) ELISA quantification of MYOM3 before the escape test (average ± SD, n = 5). ( D ) Global force evaluation (normalized to mice body weight, N/g) by the escape test (average ± SD, n = 5). ( E ) Histological characterization of muscles. Top panel: H&E labeling of GA muscle cross sections. Middle panel: Sirius red labeling of diaphragm and GA muscles. Bottom panel: Representative images of GA muscle serial cross sections immunostained for mouse immunoglobulin (IgG), laminin, and CD11b. Scale bars, 200 μm (H&E) and 500 and 100 μm (zoomed-in images). ( F ) Analysis of myofiber size distribution in GA muscles represented by the variance coefficient calculated as [SD of the muscle fiber size/mean of muscle fiber size] * 1000 (median with min-max values). ( G ) Centronucleation index in the GA muscle (median with min-max values). ( H and I ) Fibrosis analysis of GA and diaphragm by the quantification of Sirius red–positive areas on transversal sections (average ± SD). ( J ) Quantification of IgG uptake by myofibers (positive myofiber number normalized to muscle cross-sectional area). ( K ) Evaluation of CD11b + cell infiltration in the muscle by the quantification of CD11b area. An ANOVA test with Tukey’s multiple comparisons test was used for statistical comparisons (compared to WT control and mdx control). ANOVA: * P < 0.05, ** P < 0.05, *** P < 0.001, and **** P < 0.0001. Black asterisks represent relative comparison to the “WT control” group, and red asterisks represent comparison to the “mdx control” group. An unpaired two-tailed t test was used for statistical comparisons between mdx-μDys and mdx_μDys + TR groups. t test: * P < 0.05 and ** P < 0.01.

Article Snippet: An anti-MYOM3 polyclonal capture antibody (Proteintech Lab, ref. 17692-1-AP) was coated to the wells of a multiarray plate containing electrodes (MSD, ref. L15XA-3) (overnight, 4°C).

Techniques: Injection, Sequencing, Enzyme-linked Immunosorbent Assay, Muscles, Labeling, Control, Comparison, Two Tailed Test