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Figure 5 | BMC Molecular Biology

Figure 5

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

Figure 5

APOB skip 27 ASOs have no effect on the alternative splicing of other exons. (a) The indicated ASOs were transfected at 250 nM into HepG2 cells, and total RNA from these was analyzed by RT-PCR for inclusion/skipping of APOB exon 27; GAPDH, as loading control; inclusion/skipping of insulin receptor (INSR) exon 11; inclusion/skipping of TSC2 exon 25; and inclusion/skipping of Caspase-9 exons 3–6. The identity of each band is listed below each set of 6 lanes from top band to bottom band: APOB inc 27 = exon 27 included; APOB skip 27 = exon 27 skipped, GAPDH = amplified region of GAPDH mRNA 7–261 nt; INSR B = exon 11 included, INSR A = exon 11 skipped; TSC2 +25 = exon 25 included, TSC2 -25 = exon 25 skipped; Caspase-9 isoform 9 = exons 3–6 included, Caspase-9 isoform 9b = exons 3–6 skipped. Transfection of ASOs does not affect the alternative splicing of INSR, TSC2 and Caspase-9. (b) Quantitative data for alternative splicing of each gene from three independent replicates. Error bars represent S.E.M. One-way repeated measures ANOVA of the APOB skip 27 dataset showed p = 0.0007, representing significant statistical differences between groups: Dunnett's multiple comparison test showed statistically significant differences between the no-oligo control and skip 27–53, 5B and 3B, as indicated. One-way ANOVA p-values for the INSR, TSC2 and Caspase-9 datasets were 0.3716, 0.5887, and 0.1674 respectively, indicating no statistically significant differences between groups.

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