Ubiquilin-1 immunoreactivity is concentrated on Hirano bodies and dystrophic neurites in Alzheimer's disease brains
Version of Record online: 28 OCT 2013
© 2013 British Neuropathological Society
Neuropathology and Applied Neurobiology
Volume 39, Issue 7, pages 817–830, December 2013
How to Cite
Satoh, J., Tabunoki, H., Ishida, T., Saito, Y. and Arima, K. (2013), Ubiquilin-1 immunoreactivity is concentrated on Hirano bodies and dystrophic neurites in Alzheimer's disease brains. Neuropathology and Applied Neurobiology, 39: 817–830. doi: 10.1111/nan.12036
- Issue online: 28 OCT 2013
- Version of Record online: 28 OCT 2013
- Accepted manuscript online: 20 FEB 2013 05:38AM EST
- Manuscript Accepted: 7 FEB 2013
- Manuscript Received: 29 AUG 2012
- Research on Intractable Diseases. Grant Numbers: H21-Nanchi-Ippan-201, H22-Nanchi-Ippan-136
- Ministry of Health, Labour and Welfare (MHLW), Japan
- Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan. Grant Numbers: S0801043, C22500322
Figure S1. Amino acid sequence similarity between human ubiquilin-1 and ubiquilin-2 proteins. Amino acid sequences of human ubiquilin-1 and ubiquilin-2 proteins are aligned by using CLC Free Workbench version 4.5.1 (CLC Bio, Aarhus, Denmark). They showed an amino acid sequence identity of 84.9%. A mouse monoclonal antibody 5F5 is raised against the human ubiquilin-2 C-terminal peptide spanning amino acid residues 555–624 highlighted by red line.
Figure S2. Ubiquilin-1-immunoreactive Hirano bodies in the hippocampal CA1 region of non-AD brains. The tissue sections of the hippocampal CA1 region of AD, ALS, PD and normal control (NC) cases were processed for immunohistochemistry. The panels (a–d) represent (a) NC, ubiquilin-1, (b) ALS, ubiquilin-1, (c) PD, ubiquilin-1 and (d) AD, cofilin-1. Both Hirano bodies and GVD granules express cofilin-1.
Figure S3. Immunoprecipitation (IP) analysis of the interaction between ubiquilin-1 and ubiquilin-2. The full-length ORFs of ubiquilin-1 and C9orf72 cloned in the vectors expressing EYFP-tagged or DsRed-tagged proteins were coexpressed in HEK293 cells. The cell lysate was immunoprecipitated with anti-ubiqulin-1 antibody (sc-14652) or an equivalent amount of normal goat IgG, or immunoprecipitated with anti-C9orf72 antibody (sc-138763) or an equivalent amount of normal rabbit IgG. It was processed for Western blot (WB) with anti-C9orf72 antibody or anti-GFP antibody capable of detecting EYFP-tagged proteins. The panels (a,b) represent (a) IP with anti-ubiquilin-1 antibody followed by WB with anti-C9orf72 antibody and (b) IP with anti-C9orf72 antibody followed by WB with anti-GFP antibody. The lanes (1–3) indicate (1) input control, (2) IP with specific antibodies and (3) IP with normal IgG.
Table S1. Demographic profile of the cases examined in the present study.
Table S2. Primary antibodies utilized for immunohistochemistry.
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