Research article
Expression of mutant TDP-43 induces neuronal dysfunction in transgenic mice
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* Corresponding author: Leonard Petrucelli petrucelli.leonard@mayo.edu
1 Department of Neuroscience, Mayo Clinic, (4500 San Pablo Road), Jacksonville, (32224), USA
2 Center for Translational Research in Neurodegenerative Disease (CTRND), College of Medicine, University of Florida, (1275 Center Drive), Gainesville, (32610), USA
3 Department of Neuroscience, College of Medicine, University of Florida, (1275 Center Drive), Gainesville, (32610), USA
Molecular Neurodegeneration 2011, 6:73 doi:10.1186/1750-1326-6-73
Published: 26 October 2011Additional files
Additional file 1:
Figure A1: Increased ubiquitin levels in TDP-43M337V mice but hTDP-43 itself is not ubiquitinated. Human TDP-43 was immunoprecipitated from brain homogenates derived from nontransgenic and homozygous TDP-43m337v mice. Briefly, brain homogenates containing 500 μg protein were incubated with 1.5 μg mouse monoclonal TDP-43 antibody overnight at 4°C with gentle shaking. Protein G agarose was added for 4 h at 4°C then pelleted by centrifugation. Protein thus captured was eluted using sample loading buffer and resolved by SDS/PAGE for Western blot analysis. Shown are immunoblots of the inputs and the immunoprecipitated proteins, probed using an antibody to ubiquitin or to total TDP-43. Note that a marked increase in ubiquitin levels is observed in the homogenates derived from homozygous mice prior to immunoprecipitation. Nonetheless, the immunoprecipitated human TDP-43 is not immunopositive for ubiquitin. Arrow = IgG Heavy Chain.
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Additional file 2:
Figure A2: No mitochondrial fission and fusion protein changes in TDP-43M337V mice. Immunoblot analysis of Ser616-phosphorylated DLP1, DLP1, Fis1, and mitofusin 1 (MFN1) expression level in brain lysates of nontransgenic(NT), hemizygous(Hemi), and homozygous(Homo) TDP-43M337V mice of both line 4 and line 6. There are no significant protein changes among different mice groups.
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Additional file 3:
Additional Table: Primary Antibody List. Full list of the primary antibodies used in this study.
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