In addition, Western blot protein analyses demonstrated that the levels of SMN protein significantly increased in the cells by transfection of AONs #1–5 (a 1.5–1.9-fold increase from the control, Fig. 4). The AONs #6–8 treatment did not change the levels of SMN protein in the cells. These data indicate that the LNA/DNA mixmers targeting ISS-N1 in SMN2 intron 7 induce the exon 7 inclusion efficiently in the SMA patient cells.Figure 4Transfection of the novel LNA/DNA mixmers increases the production of SMN protein. (a) Western blotting of SMN and Cofilin of healthy and AON-treated SMA patient fibroblasts (GM03813). (b) Fold increase in SMN expression of AON-treated SMA patient fibroblasts. 5 nM transfection of AON 1–5 significantly increased the production of SMN proteins. Cofilin was used as a loading control. The data was normalized to the ratio of SMN/Cofilin in non-treated SMA fibroblasts. Bars represent mean ± S.D. of four independent experiments. One-way ANOVA with Dunnett’s multiple comparison test. M: mock control AON, NT: non-treated.