The SORLA coding sequence was inserted into pDEST-ORF-V1 (#73637, Addgene) and pDEST-ORF-V2 (#73638, Addgene) vectors already containing the two Venus fragments (V1 and V2) obtaining the SORLA-V1 and SORLA-V2 constructs as previously described (33 (link)).
The SORLA expression constructs including untagged SORLA (from human cDNA), SORLA-GFP, and SORLA-FLAG were cloned into the pcDNA3.1 vector as previously described in detail (47 (link)). The APP-myc construct was generated from human cDNA and introduced in the pcDNA3.1 vector followed by insertion of a C-terminal myc-tag as explained previously (48 (link)). For the recombinant expression of SORLA VPS10p-domain, the coding sequence corresponding to 1 to 753 residues of human SORLA was fused with PA-tag coding sequence on its C terminus and inserted into pcDNA3.1 in a same way previously described (40 (link)).
The SORLA expression constructs including untagged SORLA (from human cDNA), SORLA-GFP, and SORLA-FLAG were cloned into the pcDNA3.1 vector as previously described in detail (47 (link)). The APP-myc construct was generated from human cDNA and introduced in the pcDNA3.1 vector followed by insertion of a C-terminal myc-tag as explained previously (48 (link)). For the recombinant expression of SORLA VPS10p-domain, the coding sequence corresponding to 1 to 753 residues of human SORLA was fused with PA-tag coding sequence on its C terminus and inserted into pcDNA3.1 in a same way previously described (40 (link)).
