Figure 6.

Effect of ASO 7–26 in patient fibroblasts. (A) Exon-20 splicing patterns of endogenous IKBKAP mRNA were measured by RT-PCR in patient-derived GM04899 skin fibroblasts treated with 100 μM kinetin (right panel) or transfected with increasing doses of ASO 7–26 (left panel); RNA was extracted 3 days after the treatments. The IKBKAP mRNA level in the normal fibroblast cell line IMR-90 was used as reference. (B) Western blot showing IKAP levels after treatment with ASO 7–26 (left panel) or kinetin (right panel). (A) and (B), multiple lanes for each condition represent independent experiments. (C) Quantification of exon 20 % inclusion, based on the data shown in (A) and additional experiments (n = 3 independent transfections, ***P< 0.001 versus NT, one-way ANOVA). (D) Quantification of (B) showing IKAP levels normalized to β-actin. The IKAP levels in cells treated with ASO 7–26 were normalized to that of control-ASO-treated cells (n = 3 independent transfections, ***P< 0.001 versus CTRL ASO, Student's t-test); the IKAP levels of kinetin-treated cells were normalized to that of solvent-treated cells (n = 3 independent treatments, n.s. P> 0.05 versus solvent-treated, Student's t-test). In (C) and (D), NT = no-treatment, CTRL ASO = unrelated negative control ASO, and Kn = kinetin treatment. Error bars indicate standard deviation.