Extended Data Fig. 1: Intraventricular ASO delivery efficiently reduces stathmin-2 expression in mouse cortex and spinal cord.
(a-b) Stmn2 mRNA levels detected by FISH (a), and immunofluorescence confocal image immunolabeled for stathmin-2 protein (b), from 12-month-old WT mice spinal cord hemisections. Green arrows: α-motor neurons; blue arrows: γ-motor neurons; red arrows: interneurons. (c) Immunofluorescence confocal image of 12-month-old WT mice gastrocnemius muscle revealing stathmin-2 presence at the neuromuscular junction. (a-c) At least n=3 animals per condition were imaged with similar results. (d,e) Quantification of Stmn2 mRNA levels by qPCR (d) and immunoblots (e) showing stathmin-2 protein levels in mice cortex 2 weeks after the ICV injection of non-targeting (n=4 animals) or Stmn2 targeting ASOs (n=2 animals/per ASO). HSP-90 was used as a loading control in the immunoblotting. Statistics by two-sided, one-way ANOVA Dunnett’s multiple comparison test (P < 0.0001). (f) Immunoblots showing stathmin-2 protein levels in mouse spinal cord 2 weeks after the ICV injection of non-targeting or Stmn2 targeting ASOs. HSP-90 was used as a loading control. *Indicates non-specific band. (g) Quantification of Stmn2 mRNA levels by qPCR in mouse spinal cord 8 weeks after the ICV injection of non-targeting (n=4 animals) or Stmn2 targeting ASOs (n=4 animals/per ASO). Gapdh was used as an endogenous control gene. *** P = 0.0002 and **** P < 0.0001 (h,i) Quantification of Stmn2 mRNA levels by qPCR, (P < 0.0001) (h), and immunoblots (i) showing stathmin-2 protein levels in mice cortex 8 weeks after the ICV injection of non-targeting (n=4 animals) or Stmn2 targeting ASOs (n=4 animals/per ASO). HSP-90 was used as a loading control in the immunoblotting. Gapdh was used as an endogenous control gene. (j) Compound muscle action potential (CMAP) measurements in muscles of WT mice treated with non-targeting (n=4 animals) or Stmn2 targeting ASOs (n=4 animals/per ASO) for 8 weeks. (g,h,j) Statistics by two-sided, one-way ANOVA Dunnett’s multiple comparison test. All panels: Each data point represents an individual mouse. Error bars plotted as SEM. ****, P <0.0001; ***, P < 0.001; **, P < 0.01; *, P <0.05; ns, P >0.05.
