Nanobit system vectors

All polymerase chain reaction amplification and site-directed mutagenesis, including point and deletion mutations, were performed using SuperFi DNA polymerase (Thermo Fisher Scientific). Subcloning of open reading frames and their derivatives into expression plasmids, including pICE (a gift from Steve Jackson, Addgene plasmid #46960), pGEX-4T-1 (GE Healthcare Life Sciences), pLenti.puro (a gift from Melina Fan, Addgene plasmid #74218), and NanoBiT system vectors (Promega), was performed using appropriate restriction enzyme sites. In addition, the LentiCRISPR v2 vector (a gift from Feng Zhang, Addgene plasmid #52961) was used for the lentiviral transduction of GABARAPL1 sgRNA into HeLa.Kyoto, iSLK.BAC16, iBCBL-1 cells, and NIX sgRNA into iBCBL-1 cells.

All polymerase chain reaction (PCR) amplification and site-directed mutagenesis, including point and deletion mutations were performed using Platinum™ Pfx or SuperFi™ DNA polymerase (Thermo Fisher Scientific). Subcloning of open reading frames (ORFs) and their derivatives into expression plasmids including pICE (a gift from Steve Jackson; Addgene plasmid #46960), pGEX-4T-1 (GE Healthcare Life Sciences), and NanoBiT® system vectors (Promega) was performed using appropriate restriction enzyme sites (Supplementary Table 1). Overlap extension PCR^{56 (link)} was carried out for replacement of the NIX tail-anchor (TA) region with the TA region of other tail-anchored proteins and the fused genes were cloned into vectors pBiT1.1-N (Promega) and pICE_V5^{33 (link)}. Primers are listed in Supplementary Table 3.