1. Fig. 4. Analysis of brain and spinal cord of treated Gaa−/− mice and controls.
Analysis of brain and spinal cord of treated Gaa−/− mice and controls. (A to I) Four-month-old mice treated with PBS or AAV8 vectors (2 × 1012 vg/kg) and followed for 3 months (n = 4 to 5 per cohort) or 10 months (n = 8 to 9 per cohort; n = 3 Gaa−/−-PBS). Gaa+/+-PBS, wild-type littermates; Gaa−/−-PBS, untreated control. Western blot analysis of brain (A) and cervical spinal cord (B) lysates 10 months after treatment using a monoclonal anti-GAA antibody. An anti-tubulin antibody was used as loading control. (C) Quantification of glycogen content in brain 3 and 10 months after treatment. The quantification is reported as percentage of glycogen in Gaa−/− mice treated with PBS. (D to I) Analysis of spinal cord 10 months after treatment. (D) Count of choline acetyl transferase–positive (ChAT+) motor neurons (MN) in spinal cord. (E) Representative images of ChAT staining. (F) Count of ionized calcium binding adaptor molecule 1–positive (Iba1+) cells in the gray matter of spinal cord. (G) Representative images of Iba1 staining. (H) Glial fibrillary acidic protein (GFAP) fluorescence quantification in the gray matter of the spinal cord. AU, arbitrary units. (I) Representative images of GFAP staining. In all images, cells were stained with the nuclear marker 4’,6-diamidino-2-phenylindole (DAPI). Scale bars, 200 μm (I, top) and 25 μm (E, G, and I, bottom). (D to I) Gaa−/−-PBS, n = 2; n = 3 for the other cohorts. Error bars represent SD of the mean. Statistical analysis: (C) two-way ANOVA with Tukey’s post hoc (treatment, time); (D, F, and H) two-way ANOVA with Tukey’s post hoc (treatment, region of spinal cord).
Francesco Puzzo et al., Sci Transl Med 2017;9:eaam6375
Published by AAAS