Figure 3

Dose titration of selected APOB skip 27 ASOs. (a) HepG2 cells were transfected with the indicated ASOs at increasing concentrations from 25 to 250 nM for 48 hours. RT-PCR was carried out on the total RNA extracted from these cells with oligonucleotides annealing to the adjacent exons 26 and 28. The -RT control PCR was performed without reverse transcriptase. The positions of the bands corresponding to the APOB mRNA species with exon 27 (inc 27) and without exon 27 (skip 27) are indicated on the right side. Fragment lengths in bp of pBR322-MspI markers are indicated on the left side. (b) The graph shows the quantitative data from three independent replications of the experiment (error bars show the S.E.M.). The data was analyzed using a repeated measures two-way ANOVA, using Bonferroni's test to compare exon 27 skipping between skip 27-3B and the other ASOs at 250 nM: statistically significant differences are seen with skip 27–53, skip 27-3B scram and skip 27-3B A1. No statistically significant difference is seen with skip 27-5B and skip 27-3B mutA1.