Extended Data Fig. 7: ALS-linked mutation in FUS does not impair hypusine levels nor total protein synthesis in cell bodies.
From: Axonal Eif5a hypusination controls local translation and mitigates defects in FUS-ALS
a, Immunofluorescence analysis of puromycin, hypusine and 4′,6-diamidino-2-phenylindole (DAPI) in nontransgenic or hFUSR521H-derived primary cortical neuron cell bodies. Scale bar, 10 µm. b,c, Quantifications of hypusine (b) and puromycin (c) signals in the indicated conditions. Data are represented as mean ± s.e.m. (n = 3 biological replicates). P values are calculated using one-way ANOVA with the Bonferroni multiple comparisons test. d, Immunofluorescence analysis of puromycin, hypusine and DAPI staining in nontransgenic or hFUSR521H-derived primary motor neuron cell bodies, treated in the axonal compartment with spermidine 100 µM for 48 h and pulse-labeled with puromycin. Scale bar, 10 µm. e,f, Quantifications of hypusine (e) and puromycin (f) signals in the indicated conditions. Data are represented as mean ± s.e.m. (n = 3 biological replicates). P values are calculated using one-way ANOVA with the Bonferroni multiple comparisons test. g, Axonal branching quantification from Fig. 6e. Data are represented as mean ± s.e.m. (n = 3–5 biological replicates). P values are calculated using one-way ANOVA with the Bonferroni multiple comparisons test. h, Effect of spermidine treatment ranging between 0 and 5 mM on the eclosion of the control fly line. Data are represented as mean ± s.e.m. (n = 15–16 crosses/condition from three independent experiments). P values are calculated using one-way ANOVA with Holm–ŠÃdák’s multiple comparisons test.
