Extended Data Fig. 5: Slc33a1 loss promotes UPR transcriptional signatures and GSH synthesis inhibitors rescue Slc33a1-dependency.
From: Keap1 mutation renders lung adenocarcinomas dependent on Slc33a1
a-d, GSEA enrichment plots of conserved ER stress response signatures (ER chaperones FDR = 0.0; Aminoacyl tRNA synthetase FDR = 0.0; ER/Golgi traffic FDR = 0.0146; ERAD FDR = 0.025). Analysis derived from a single experiment from KP-sgCtrl, KP-sgSlc33a1, KPK-sgCtrl, KPK-sgSlc33a1 samples. e, GSH concentrations measured in cells treated with or without 48 hours of (25 μM) BSO or (780 nM) Erastin. Data are representative of n = 3 independent culture wells per cell line per sgRNA. f, Viability of cells infected with sgRNAs and treated with or without 48 hours of BSO and Erastin as in (e). Viability was measured using cell-titer Glo. Data are representative of n = 3 independent culture wells per cell line per sgRNA per treatment condition. g, Colony formation assay of KPK1 cells transduced with LVt-TSTOP vectors expressing shRenilla or shSlc33a1. h, Day 2 normalized fluorescence competition assays of 50 μM BSO or 1 μM Erastin treated pUSEPR-infected KP or KPK isogenic cell lines expressing sgCtrl or sgRpa3. Data are representative of n = 3 independent culture wells per cell line per sgRNA. i, Total cell counts of KPK-Slc33a1 knockout cells treated with indicated conditions 7 days post infection. Data are representative of n = 3 independent experiments. P values were determined by ANOVA with Dunnet’s multiple comparisons testing. All error bars depicted represent mean ± s.d. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, n.s. = not significant. Data from a single experiment are shown in e, f, g, and h are representative of two independent experiments with similar results. Data for experiments e, f, h and i are available as source data (Source_Data_Extended_Data_1; Source_Data_Extended_Data_3).
