Cell Line Overexpression and Knockdown

HA‐LIPIN1^WT, HA‐LIPIN^Mut, and mitoPLD‐GFP overexpression constructs were kind gifts from Hiromi Sesaki (Johns Hopkins) and described previously (Adachi et al, 2016 (link)). The PA‐PLA1‐GFP overexpression construct was purchased from Addgene (#162880). Perk^WT and Perk^PSP overexpression plasmids were kind gifts from Jonathan Lin (Stanford; Yuan et al, 2018 (link)). Plasmids containing shRNA were purchased from Sigma in the pLKO.1 vector: mouse Prelid1 shRNA (TRCN0000345802), human PRELID1 shRNA (TRCN0000130829). All compounds used in this study were purchased: thapsigargin (Tg; Fisher Scientific), GSK2656157 (BioVision Inc.), ISRIB (Sigma), CCCP (Sigma), rapamycin (Selleckchem), and ionomycin (Sigma).