Figure 4.

Comparison of AON efficiency in mutant and wild-type COCH-minigene-expressing cells

(A and B) AONs directed against the c.151C>T mutation or the c.436+368_436+369dupAG (dupAG) variant were transfected in stable transgenic cell lines expressing (A) a mutant COCH minigene, and (B) a wild-type COCH minigene. AONs were transfected at a dose that results in a final concentration of 100 nM in the culture medium, and their effect on COCH transcript levels was investigated 24 h post transfection. (A) As shown previously, c.151C>T AON-B and AON-E, and dupAG AON-B, were able to induce a significant decrease in the mutant COCH transcript level. (B) None of the AONs resulted in a significant decrease in wild-type COCH transcript levels in transgenic cells expressing the wild-type COCH minigene. While c.151C>T AON-A results in a decrease in wild-type COCH transcript levels, the observed decrease is not statistically significant (p = 0.14, Tukey’s multiple comparison test). All AONs used here consisted of a gapmer composition. Mutant and wild-type COCH transcript levels are normalized for the expression of RPS18 and plotted as mean ± SD of 3 replicates. The left axis shows the fold change compared to scrambled AON-transfected cells; the right axis shows the RPS18 normalized transcript levels. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, one-way ANOVA with Tukey’s post-test.