FIG. 3.

Test of AONs and lentiviral vectors to skip the 101bis pseudo-exon. (a) Seven AONs (A to G) were tested on MF cells for their capacity to skip the pseudo-exon. MF cells were either not transfected (lane MF) or transfected with each of the AONs (A to G) or with pairs of AONs (D+E or E+G). Cells were then induced to differentiate for 4 days and RT-PCR was performed using same primers as in Fig. 2a. (b) Ratios of the amount of pseudo-exon-containing fragment versus the total amount of amplified fragments (normal+pseudo-exon containing fragments) were calculated for each condition and normalized to 1 for nontransfected MF cells. Results are given as mean±SEM of three experiments. (c) Lentiviral U7-snRNA vectors containing antisense sequences corresponding to a nonrelevant control AON (U7-Ctrl) or to AONs D+E (U7-D+E) were tested for their efficacy to skip the pseudo-exon. MF cells were either not transduced (lane MF), or transduced with U7-Ctrl or U7-D+E vectors. Cells were then induced to differentiate for 4 days and RT-PCR was performed as in (a). (d) Quantifications were performed as in (b), and the mean±SEM of four experiments is plotted. ***p<0.001, Student's t-test comparison of MF and MF U7-D+E cells. AONs, antisense oligonucleotides; snRNA, small nuclear RNA.