Fig. 2.

ASO-10-27 treatment boosts SMN protein expression in SMA mice. Treatment with ASO-10-27 increased exon 7 inclusion during SMN2 splicing, and increased SMN protein expression and the numbers of motor neurons in the spinal cord after intracerebroventricular injection of neonatal mice with severe SMA at postnatal day 0 (P0). (A) The amount of SMN2 mRNA that contained exon 7 relative to SMN2 mRNA in untreated SMA mice was determined by reverse transcription–polymerase chain reaction (RT-PCR) at 16 days. (B) The amount of SMN protein relative to that in untreated wild-type (WT) mice was determined by Western blotting at 16 days. (C) For the pharmacodynamic/pharmacokinetic analysis, the absolute ASO tissue concentration in the cervical region was determined by capillary gel electrophoresis; the amount of SMN protein relative to that in untreated WT mice in the thoracic region was determined by Western blotting; and the amount of SMN2 mRNA that contained exon 7 relative to that in untreated SMA mice in the lumbar region was determined by RT-PCR at 3, 16, and 30 days. (D) The number of motor neurons in the cervical, thoracic, and lumbar regions was determined by ChAT immuno-staining at 16 days. SMA, untreated SMA mice; MM, ASO-mismatch–treated SMA mice; 10-27, ASO-10-27–treated SMA mice; WT, untreated WT mice. The P values between the different treatment groups were determined by one-way ANOVA and Bonferroni multiple post hoc comparisons (P < 0.01; P < 0.001). Data are means ± SEM.