Figure 3
Alternative splicing of SMA target genes is altered in hFUS+/+ mice. (a) Alternative splicing pattern of selected SMA target exons in hFUS+/+ and hFUS−/− mice. Spinal cords from two different end stage FUS+/+ mice were analysed, together with age-matched control animals. Results are representative of at least n = 3 independent experiments with 5 different mice. The exons analysed for each gene are reported in Supplemental Table 3. For Gria4, Vps16, and Agrin, different exons were examined, as indicated in subscripts. Schematics of exons (rectangles) and introns (lines) analysed are shown on the left. Full-length agarose gels are presented in Supplementary Figure 7. (b) Bands from the experiments exemplified in Fig. 3a were quantified by densitometry analysis, and a splicing index was calculated as follows: for genes that are expressed in more than one isoform, depending on the splicing event involved (i.e. exon skipping/intron retention), the ratios between the upper and the lower bands were calculated and scaled to have the average ratio in hFUS−/− mice at 1. For the Agrin gene (exon 31–34), the upper (u), central (c) and lower (d) bands correspond to isoforms containing exon 31/32/33/34, 31/33/34 and 31/34, respectively. The ratio between bands c/d, u/d and uc/d were calculated as indicated. For genes that are expressed as a unique isoform, the splicing index was calculated as the ratio between band intensity and the relative intensity of the housekeeping Gapdh mRNA. Results from n = 5 independent mice have been considered for each genotype. Mean ± SD is shown. One asterisk is shown when p ≤ 0.05, two asterisks when p ≤ 0.01.
