Figure 6
SMA molecular phenotypes are not affected by SMN depletion. (a) Quantitative analysis of Cresyl Violet stained motor neurons in the ventral horns of spinal cord from hFUS+/+; Smn+/+ and hFUS+/+; Smn+/− mice at 40 days. n = 4 animals were used for each genotype. Values are reported as mean ± SD. (b) Alternative splicing patterns of the indicated target genes in endstage and control mice with the indicated genotypes. Results are representative of at least n = 3 independent experiments. Full-length agarose gels are presented in Supplementary Figure 9. (c) The number of Y12-positive snRNPs were counted in the motor neurons of hFUS+/+; Smn+/+ and hFUS+/+; Smn+/− mice at 40 days. Results are reported as means ± SD. A total number of 50 motor neurons were scored. (d) RT-qPCR quantification of snRNA U1 and U2 in lysates from the spinal cords of the indicated mice. Means ± SE were normalized for the snRNA levels in control hFUS−/− mice. (e) Expression levels of Gemin2, Sm B/B’, SMN, FUS and β-actin were assayed by Western blot on spinal cord lysates from 40 days-old mice of the indicated genotypes. The panel is representative of at least n = 3 independent experiments. Full-length blots are presented in Supplementary Figure 10. (f) The expression levels of SMN, Gemin2 and Sm B/B’ in hFUS+/+ mice were calculated by densitometric analysis of the bands from Western blots as in (e), normalised to β-actin levels and expressed as fold increases over the hFUS−/−; Smn+/+ control animals. Means ± SD are shown from n = 3 independent experiments. Values significantly different from relative controls are indicated with an asterisk when p ≤ 0.05.
