FIG 1.

Development of an assay to identify compounds which are synthetic lethal with Nrf2. (A) Scheme showing an overview of the synthetic lethal screening strategy using isogenic WT and Keap1 KO Hepa1 cells. (B) Nrf2 target genes are significantly upregulated in CRISPR-Cas9-generated Keap1 KO Hepa1 cells compared to the parental WT cells. (C) WT-GFP cells plated at multiple densities display normal growth dynamics over a 5-day period. (D) Keap1 KO-mCherry cells plated at multiple densities display normal growth dynamics over a 5-day period. (E) When cocultured together at an initial seeding of 1,000 WT-GFP cells and 2,000 Keap1 KO-mCherry cells, the cell lines proliferate together at an expected rate over an 8-day period. (F) Over a 4-day period, the fluorescence intensity of the WT-GFP and Keap1 KO-mCherry monocultured cells increases at a rate comparable to the increase in total protein content. (G) Visualization of the coculture of WT-GFP and Keap1 KO-mCherry cells, showing uniform fluorophore expression between the cells. Scale bars = 300 μm. (H and I) Under coculture conditions, compared to the Keap1 KO-mCherry cells, the WT-GFP cells are significantly more sensitive to the anticancer drugs 5-FU and doxorubicin (Dox).