Fig. 2: Dose-dependent toxicity and detergent insolubility of mutant FUS. | Nature Medicine

Fig. 2: Dose-dependent toxicity and detergent insolubility of mutant FUS.

From: Antisense oligonucleotide silencing of FUS expression as a therapeutic approach in amyotrophic lateral sclerosis

Fig. 2

a, Kaplan–Meier survival curves for mice with the indicated combinations of Fus WT, P517L, Δ14 and null-knockout alleles. b, Median survival of selected genotypes plotted on logarithmic scale illustrates partial functionality and dose-dependent toxicity of mutant FUS. Increased survival of P517L/KO and Δ14/KO compared with KO/KO animals shows that mutant FUS protein is able to partially rescue the null phenotype and, thus, is functional. Comparison of P517L/P517L and P517L/Δ14 versus P517L/KO animals shows that further addition of mutant FUS protein decreases survival, consistent with dose-dependent toxicity of mutant FUS protein. Median survival was estimated for P517L/P517L (0.15 d) and KO/KO (0.2 d), as most newborn pups for these genotypes were found dead and, thus, could not be accurately quantified. The bar for P517L/KO is reproduced three times for comparison purposes. c, Immunoblot of sarkosyl-insoluble fractions from brains of newborn Fus WT/WT, heterozygous P517L/WT and homozygous P517L/P517L mice. Each lane corresponds to a separate animal. d, Immunoblot of sarkosyl-insoluble fractions from brains of 2-year-old Fus WT/WT and heterozygous P517L/WT and Δ14/WT mice. Each lane corresponds to a separate animal. e, Immunoblot of sarkosyl solubility fractionation of human brain stems of a non-ALS control and a patient with ALS-FUS P525L. f, Immunoblot of sarkosyl-insoluble fractions from human brain stem samples from a non-ALS control and a patient with ALS-FUS P525L. g, Relative abundance of inclusion isoforms of exons regulated by hnRNPH. The observed pattern is consistent with functional deficiency of hnRNPH, which promotes inclusion of exon 24 of ATXN2 (left) and inhibits the inclusion of exon 8 of hnRNPDL transcript (right). *P < 0.05, **P < 0.01 and ***P < 0.001, using one-way ANOVA with Tukey’s post hoc test. n = 4 animals per genotype. SI, sarkosyl insoluble; Sol, soluble (in hypotonic buffer); SS, sarkosyl soluble (in 1% sarkosyl and high salt).

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