Figure 6

Antisense oligonucleotides directed against identified ESS elements cause alternative splicing of APOB in HepG2 cells. In lanes 1 and 2 the cells were not transfected with ASOs. The –RT control PCR (lane 1) was performed without reverse transcriptase. In lane 3, the cells were transfected with negative control ASO at 500 nM. Lanes 4–13 were transfected with the indicated ASOs at 125 nM each; the total concentration of ASO was adjusted to 500 nM with negative control ASO. Cells were incubated for 48 hours and RT-PCR was carried out on the total RNA extracted from these cells with oligonucleotides annealing to the adjacent exons 25 and 27. The positions of the bands corresponding to the APOB mRNA species with constitutive exon 26 inclusion (top), pseudo 3^′ splice site activation (middle) and skipping of exon 26 (bottom) are indicated on the right side. Fragment lengths in bp of the markers are indicated on the left side.