Figure 1.
AAV9-U1-FD treatment (i.c.v. + i.p.) rescues life span in FD mice
(A and B) Endpoint gels (A) and relative quantification histograms (B) of ELP1 splicing isoforms in the TgFD9 asymptomatic mice treated with saline (lane 1) versus treatment with AAV9-U1-FD: intracerebral ventricular (i.c.v.) injected (lane 2), intraperitoneal (i.p.) injected (lane 3), and i.c.v. + i.p. injected (lane 4) at PND-10. Data are expressed as percentage of exon 20 inclusion. DRG, dorsal root ganglia; TG, trigeminal ganglia; SC, spinal cord. In the graph, data are reported as mean ± SEM, for each pair of animals tested. The upper band of 202 bp represents the ELP1 isoform in which exon 20 is included whereas the lower band of 128 bp represents the isoform in which exon 20 is skipped. Statistical analysis was performed by two-way ANOVA with Bonferroni correction, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.
(C) qPCR quantification of ExSpeU1-FD in the TgFD9 asymptomatic mice treated with saline (lane 1) versus treatment with AAV9-U1-FD: intracerebral ventricular (i.c.v.) injected (lane 2), intraperitoneal (i.p.) injected (lane 3), and i.c.v. + i.p. injected (lane 4) at PND-10. Data (mean ± SEM) are expressed as percentage of ExSpeU1-FD relative to endogenous U1, two technical replicates on 2 mice per group were used. DRG, dorsal root ganglia. Statistical analysis was performed by two-way ANOVA with Bonferroni correction, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001; ns, not significant.
(D) Kaplan-Meier survival curves of FD mice (IkbkapΔ20/Ikbkapflox; h-TgFD9/+) and littermate controls (Ikbkapflox/+; h-TgFD9/+) treated with one intracerebral ventricular injection (i.c.v.) at PND-0 and a second intraperitoneal injection (i.p.) at PND-2 of AAV9-U1-FD (1.6 × 1011 VG/mouse) or saline only. Littermate controls + saline in black; littermate controls + AAV9-U1-FD in gray; FD + saline in orange; FD-treated with AAV9-U1-FD in blue. Statistical analysis was performed with log-rank test, ∗∗∗∗p < 0.0001, n > 15 mice for each group.