Fig. 7: FUS accumulates in synaptosomes of FusΔNLS/+ mice and alters synaptosomal levels of a subset of its targets. | Nature Communications

Fig. 7: FUS accumulates in synaptosomes of FusΔNLS/+ mice and alters synaptosomal levels of a subset of its targets.

From: Cytoplasmic FUS triggers early behavioral alterations linked to cortical neuronal hyperactivity and inhibitory synaptic defects

Fig. 7

a, b Representative western blot images (a) and respective quantifications (b) of cytoplasmic (a, left) or synaptosome (a, right) extracts from Fus+/+ (+/+) or Fus∆NLS/+ (∆/+) mice (4 months of age,) using two antibodies recognizing the N-terminal part of the FUS protein (FUS N-ter1 and FUS N-ter2), the C-terminal part of FUS (encoding the NLS, FUS C-ter) or synaptophysin protein to show enrichment in synaptic proteins in the synaptosome fraction. N = 6 Fus+/+ mice and N = 6 Fus∆NLS/+ mice were analyzed. Data are presented as mean ± SEM. One-Way ANOVA with Tukey post-hoc test. ***p < 0.0001 Please note that the FUS western blots were run on independent gels, to avoid stripping and reprobing on the same membrane for the same protein. Each of these gels were controlled for equal loading using StainFree markers, that are provided in the source data. c, d Representative western blot images (c) and respective quantifications (d) of total extracts (c) from Fus+/+ (+/+) or Fus∆NLS/+ (∆/+) mice (4 months of age,) using the same antibodies as in panel a. N = 5 Fus+/+ mice and N = 5 Fus∆NLS/+ mice were analyzed. Data are presented as mean ± SEM. Two-tailed Unpaired Student’s t-test. N-ter1: p < 0.0001; C-ter: p-value: p = 0.0006; Synaptophysin: p = 0.0411. e mRNA levels of the indicated genes in RNAs extracted from cytoplasmic (Cyto.) or synaptosome (Synap.) extracts from Fus+/+ (+/+) or Fus∆NLS/+ (∆/+) frontal cortex from 4-months-old female mice as assessed using RT-qPCR. N = 6 Fus+/+ mice and N = 5 Fus∆NLS/+ mice were analyzed. Data are presented as mean ± SEM. Genes are grouped by categories (controls, established FUS RNA targets, and genes belonging to the Turquoise module). All quantifications are presented relative to the +/+ cytoplasmic RNA levels set to 1. One-way ANOVA with Tukey post-hoc test. Fus: ***p < 0.0001 vs corresponding wild-type fraction; ###p < 0.0001 vs corresponding cytoplasmic fraction of the same genotype. Malat: ###p = 0.0001 vs corresponding cytoplasmic fraction of the same genotype. Nrxn1 *p = 0.0140 vs corresponding wild-type fraction; ###p < 0.0001 vs corresponding cytoplasmic fraction of the same genotype. Gabra1: **p = 0.0012 vs corresponding wild-type fraction; ###p < 0.0001 vs corresponding cytoplasmic fraction of the same genotype. Gabrb1: **p = 0.0029 vs corresponding wild-type fraction; ###p < 0.0001 vs corresponding cytoplasmic fraction of the same genotype. Grid2: ###p < 0.0001 vs corresponding cytoplasmic fraction of the same genotype. Ctnnd2: no significant differences observed (p > 0.05). Source data are provided as a Source Data file.

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