FBXW8 is shifted into insolubility in <i>Atxn2</i>-CAG42-KIN mice due to interaction with expanded ATXN2.

<p>(A) Pulling either with anti-ATXN2 or anti-FBXW8 antibody, ATNX2 and FBXW8 show an interaction in the cerebellum of 18-month-old <i>Atxn2</i>-CAG42-KIN mice independent of the polyQ length (experiment repeated three times for anti-ATXN2 and once with anti-FBXW8, representative images). (B) In cerebellar tissue of 18-month-old <i>Atxn2</i>-CAG42-KIN mice FBXW8 protein level is downregulated in the RIPA-soluble fraction while it is upregulated in the SDS-soluble fraction (two independent experiments each with 4 <i>Atxn2</i>^CAG1/CAG1 vs. 4 <i>Atxn2</i>^CAG42/CAG42 mice).</p

PARK2 interacts with FBXW8 and is recruited into insolubility in <i>Atxn2</i>-CAG42-KIN mice.

<p>(A) In HeLa cells overexpressing Cherry-GFP-PARK2 and FBXW8-HA, pulling with anti-FBXW8 antibody resulted in the detection of FBXW8 as well as of PARK2 in Co-IP lysates, demonstrating their interaction (experiment repeated twice, representative image). (B) PARK2 interacts with FBXW8 in Co-IP samples of <i>Atxn2</i>-CAG42-KIN mice independent of the polyQ length. Lower bands represent PARK2 protein (experiment repeated once). (C) PARK2 protein level is decreased in the RIPA-soluble fraction while it is increased in the SDS-soluble fraction (8 <i>Atxn2</i>^CAG1/CAG1 mice vs. ≥ 6 <i>Atxn2</i>^CAG42/CAG42 mice).</p

FBXW8 protein levels are dysregulated in SCA2 patient material.

<p>FBXW8 expression is upregulated at the transcript level in SCA2 patient skin fibroblasts (A; 4 CTL individuals vs. 4 SCA2 patients) as well as in SCA2 patient blood samples (B; 5 CTL individuals vs. 3 SCA2 patients). (C) At the protein level FBXW8 is decreased in the RIPA-soluble fraction while it is increased in the SDS-soluble fraction in SCA2 patient fibroblasts (4 CTL individuals vs. 4 SCA2 patients).</p

Distribution of LRR values of controls and individuals with CNV at SNP level.

<p>LRRs of all individuals were extracted and plotted. “<i>Cont</i>” indicates control individuals and “<i>CNV</i>” indicates individuals with a CNV. “*” indicates the significance level between controls and CNV. 4 SNPs, including one upstream, one downstream and two within the CNV region, were selected and LRRs were plotted for (a) intergenic reigon on chromosome 11, (b) MAP4K3 gene, (c) HLA-B gene and (d) EPHA3 gene.</p

Distribution of ATXN2 repeat sizes, represented as polyQ triplet numbers.

<p>Turkish ALS patients are represented in red and healthy controls in blue bars; numbers of the individuals having the relevant alleles are shown on the top of each bar.</p

The frequency of ATXN2 expansion sizes in ALS and controls (in the genotypic model).

a<p>Fisher’s exact test.</p

CNV call validation via LRR values.

<p>LRR values of controls and individuals with CNV for particular SNPs were extracted to check CNV calls of PennCNV tool. Average LRR values of each SNP for both groups were calculated and plotted. Consecutive SNPs in chromosome position line do not demonstrate the exact distance. The CNVs detected by PennCNV were shown by black bars. (a) Loss of an intergenic region on chromosome 11 between rs1916207 and rs554110 (chr11: 50,545,009–50,586,426), (b) gain of MAP4K3 gene between rs12151392 and rs2373530 (chr2: 39,372,016–39,428,488), (c) homozygous loss of HLA-B gene between rs9295976 and rs28367708 (chr6: 31,389,749–31,393,270), (d) loss of EPHA3 gene between rs2063589 and rs870899 (chr3: 89,485,137–89,499,861) (NCBI37/hg19).</p

Clinical characteristics of four Turkish ALS patients with ATXN2 expansions.

<p>AO: age of onset, AD: age at death. UE: upper extremity, LE: lower extremity, ULE: upper and lower extremity.</p>*<p>GWA- genotyped.</p

Association results of CNVs observed in analysis with p<0.05 (Fisher’s Exact Test).

<p>loss* Homozygous deletion.</p

Summary Statistics of CNV screening in 115 ALS patients and 106 controls.

<p>Summary Statistics of CNV screening in 115 ALS patients and 106 controls.</p