Figure 2: In vitro ARID1A/ATR synthetic lethality.
From: ATR inhibitors as a synthetic lethal therapy for tumours deficient in ARID1A
(a) Chemical structure of VX-970. (b) Western blot of ARID1A in human ARID1A isogenic HCT116 cells. (c) Image of colonies in six-well-plate clonogenic assay. HCT116 ARID1A isogenic (+/+ and −/−) cell lines were exposed to increasing concentrations of VE-821 (0, 0.1, 1 μM) or VX-970 (0, 0.01, 0.05 μM) for 14 days. (d–f) Dose–response clonogenic survival curves of HCT116 ARID1A isogenic (+/+ and −/−) cell lines exposed to increasing concentrations of VE-821 (d), VX-970 (e) and AZ-20 (f) for 14 days. Error bars represent s.d. (n=3), ANOVA P value of <0.0001, results are representative of triplicate biological experiments. (g–i) Dose–response survival curves of HCT116 ARID1A isogenic (+/+ and −/−) cell lines exposed to increasing concentrations of VE-821 (g), VX-970 (H) or AZ-20 (I) for 5 days. Cell viability was estimated by CellTitre-Glo reagent. Error bars represent s.d. (n=16), ANOVA P value of <0.001, results are representative of triplicate biological experiments. (j) Area under curve (AUC) box whisker comparison plot for human tumour cell lines exposed to VX-970 for 5 days. ARID1A wild-type tumour cell lines (n=15) were compared with ARID1A mutant cell lines (n=9). P=0.00594 median permutation test. (k). Dose–response clonogenic survival curve of mouse Arid1a isogenic ES cell lines. Experiment was performed as per (c). Error bars represent s.d. (n=3), ANOVA P value of <0.0001, results are representative of triplicate biological experiments. (l). Western blot of Arid1a protein expression in mouse ES Arid1a isogenic cells.
