Protocol detail

Generation of Protein Expression Constructs

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The pEGFP-N1 vectors containing human SOD1^wt, SOD1^A4V and G93A SOD1 were generated as described65 (link). Expression vector pCMV6-AC-GFP containing human TDP-43 and FUS was obtained from OriGene (USA). SOD1-tomato constructs were generated by replacing the GFP sequences in the GFP-tagged constructs65 (link) with the tomato red fluorescent protein. TDP-43^wt-tomato (Addgene plasmid 28205, provided by Zuoshang Xu34 (link)), mRFP-Ubiquitin (Addgene plasmid 11935, provided by Nico Dantuma66 (link)) and pmRFP-LC3 (Addgene plasmid 21075, Provided by Tamotsu Yoshimori67 (link)) constructs were acquired from Addgene (USA). The vectors describing Httex1(46Q) fusions (C-terminally) to mCherry and Cerulean were generated as described33 (link)68 (link).

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Farrawell N.E., Lambert-Smith I.A., Warraich S.T., Blair I.P., Saunders D.N., Hatters D.M, & Yerbury J.J. (2015). Distinct partitioning of ALS associated TDP-43, FUS and SOD1 mutants into cellular inclusions. Scientific Reports, 5, 13416.

Publication 2015

pCMV6-AC-GFP mRFP-Ubiquitin pmRFP-LC3

Human Plasmid Red fluorescent protein Tdp 43 Tomato Ubiquitin Vectors

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Protocol cited in 3 other protocols

Variable analysis

independent variables

SOD1^wt
SOD1^A4V
G93A SOD1
Human TDP-43
Httex1(46Q) fusions (C-terminally) to mCherry and Cerulean

dependent variables

Not explicitly mentioned

control variables

Not explicitly mentioned

controls

Positive controls: None specified
Negative controls: None specified

Annotations

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