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Figure 2 | BMC Molecular Biology

Figure 2

From: Antisense oligonucleotide-induced alternative splicing of the APOB mRNA generates a novel isoform of APOB

Figure 2

Alternative splicing of exon 27 induced by APOB exon 27 splice-site ASOs. (a) HepG2 cells were transfected with the indicated ASOs at 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. The band corresponding to skip 27 mRNA was cloned and sequenced to confirm that exon 26 is spliced to exon 28. (b) The graph shows the quantitative data from three independent replications of the experiment (error bars show the S.E.M.). Overall one-way repeated measures ANOVA p-value was < 0.0001, indicating statistically significant differences between groups. Dunnett's multiple comparison test was used to compare the no-oligo control versus ASO-transfected cells: statistically significant differences were found with skip 27–53, skip 27-5B, and skip 27-3B.

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