Plasmid Constructs for TDP-43 and STAU1 Studies

The plasmid constructs used in this study were 3XFlag-tagged STAU1^{7 (link)} or wildtype TDP-43,^{7 (link)} GFP, DsRED, and STAU1-DsRED. The pRK5-EGFP-Tau was a gift from Karen Ashe (Addgene, Plasmid #46904). The pBacMam2-DiEx-LIC-N-flag_huntingtin_full-length_Q23 (Addgene, Plasmid #111723) and pBacMam2-DiEx-LIC-C-flag_huntingtin_full-length_Q66 (Addgene, Plasmid #111750) were gifts from Cheryl Arrowsmith. Mutant TDP-43 cDNAs (TDP-43^G298S and TDP-43^A382T) were polymerase chain reaction (PCR)-amplified from cDNA of patients with ALS fibroblasts (FBs) with TDP-43 mutations (G298S #ND32947 and A382T #ND41003; Coriell Cell Repositories, Camden, NJ, USA). TDP-43^G348C, and C-terminal fragment of TDP-43 (TDP-43^CTF) constructs were generated using TDP-43 cDNA as template.^{7 (link)} TDP-43 with mutated nuclear localization signals (TDP-43ΔNLS) was PCR amplified from genomic DNA from hTDP-43ΔNLS mouse.^{12 (link)} The PCR products were cloned into pCMV-3XFlag plasmid (Agilent Technologies, Santa Clara, CA, USA). To generate a STAU1-tagged DsRED construct, the STAU1 coding sequence along with in frame GFP sequence was cloned into the pcDNA3 plasmid (Thermo Fisher Scientific). All constructs were verified by sequencing. 3XFlag is referred to as Flag in the text and figures.