Figure 2 | Scientific Reports

Figure 2

From: Using antisense oligonucleotides for the physiological modulation of the alternative splicing of NF1 exon 23a during PC12 neuronal differentiation

Figure 2

Design of a PMO to force the skipping of E23a and experimental conditions set up. (A) Schematic diagram of E23a skipping strategy using PMO-SkpE23a (in red) targeting its acceptor splice site. (B) Dose–response analysis at different PMO-SkpE23a concentrations in HEK293T cells measured by RT-PCR. (C) Time course experiment using PMO-SkpE23a in PC12 cells measured by RT-PCR. (D) Relative mRNA isoform type I (-E23a) expression normalized by the total Nf1 expression at 72 h, measured by RT-qPCR. (E) Total Nf1 expression levels at 72 h after PMO addition, measured by RT-qPCR. (F) Expression of Type II/I isoforms in PC12 cells treated with transfection agent (TA), a non-targeting control PMO and PMO-SkpE23a, 72 h after PMO addition and measured by RT-PCR. For all plots, bars indicate Mean ± SEM. NRE: Normalized Relative Expression. All supporting uncropped gels can be found in Supplementary Fig. S2. E23a splicing was quantified as the percent spliced-in (PSI) (standard deviation in parentheses).

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