Figure 2.

Increase in full-length SMN transcript and SMN protein levels after E1^MOv11 treatment. (a) RT-PCR image of endogenous SMN1/2-derived transcripts from HEK293T cells and (b) in spinal cord tissues from three SMNΔ7 mice. SMN2 and SMN1 transcripts are clearly distinguishable after reaction products of the cell culture extracts were digested with restriction enzyme DdeI. Plasmids containing the full-length or SMN▵7 products, pCI-▵7 and pCI-FL, respectively, were used in PCR reactions to generate size standards. Injection of the E1^MOv11 variant targeting the Element1 repressor increases total SMN protein in brain (c) and spinal cord (d) tissues of SMNΔ7 mouse model. Western blots (n = 5) for each treatment group were performed on tissues harvested at P7. (e) Western blot quantification. Western blot (n = 3) from brain and spinal cord tissue of five (5) ICV injected animals with E1^MO and E1^MOv11. Bar graph showing significant percent increase in SMN protein induction compared to the non-injected SMA control group. Significance was calculated using Student's t-test, where ns = nonsignificance, ****P ≤ 0.01, ***P ≤ 0.001. RT-PCR, reverse transcriptase-polymerase chain reaction; ICV, intracerebroventricular; SMA, spinal muscular atrophy; SMN, survival motor neuron; HEK, human embryonic kidney.