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. 2020 Oct 5;9:e56749. doi: 10.7554/eLife.56749

Figure 5. Combination of CB-839 and V-9302 depletes GSH and induces lethal ROS levels in GD liver cancer cells.

Figure 5.

(a) Intracellular GSH levels were measured by LC/MS-MS in SNU398 and HepG2 cells treated with indicated drugs for 48 hr, respectively. (b) ROS levels were measured using the CellROX Deep Red flow cytometry assay. (c) Western blot analysis was performed for γH2AX as a DNA damage marker. HSP90 served as a control. (d, e) Long-term colony formation assay and CellTiter-Blue viability assay show rescued proliferation and viability of SNU398 and HepG2 cells after ROS scavenger N-acetyl-cysteine (NAC) treatment. (f) Caspase-3/7 positive percentages of control, NAC, V-9302, CB-839, the combination, or combination plus NAC treated SNU398 and HepG2 cells in the presence of a caspase-3/7 activatable dye. (g) Schematic showing how the combination of CB-839 and V-9302 decreases GSH and induce apoptosis in liver cancer. All the data in this figure are represented as mean ± SEM. Statistical significance was assessed using a Student’s t test. *p<0.05, **p<0.01, ***p<0.001.

Figure 5—source data 1. Combination of CB-839 and V-9302 depletes GSH and induces lethal ROS level in GD liver cancer cells.