Figure 1: RNAi screen reveals genetic determinants of ATRi sensitivity.
From: ATR inhibitors as a synthetic lethal therapy for tumours deficient in ARID1A
(a) Structure of VE-821 and schematic representation describing workflow for parallel VE-821 chemosensitization screens in MCF12A and HCC1143 cells. (b) Scatter plots of VE-821 Drug Effect (DE) Z-scores from all 1280 siRNAs SMARTpools used in the chemosensitization screens. The DE Z-score threshold of −2 (dotted red line) was used for defining candidate/unvalidated synthetic lethal interactions. (c) Venn diagram of DE<−2 VE-821 sensitization hits in MCF12A and HCC1143 cells. Numbers shown indicate number of sensitization genes. Amongst the 30 genes with DE Z-scores <−2 in both cell lines, seven well-established tumour suppressor genes were identified. (d) Bar charts illustrating DE Z-scores for control, non-targeting, siRNAs (siALLSTAR, siCon1, siCon2) and ARID1A SMARTPool siRNAs in the chemosensitization screens. Values shown are medians from triplicate screens. Error bars represent s.d. (e) Three-hundred eighty-four-well plate cell survival data from HCC1143 cells transfected with siRNA targeting ARID1A (red) or siCon (blue). Twenty four hours after transfection, cells were exposed to VE-821 for 5 continuous days. Error bars represent s.d. (n=16) and results are representative of three biological replicates. Survival curve siARID1A versus siCon P value <0.0001, ANOVA. (f) Western blot illustrating ARID1A protein silencing from experiment (e). (g) Bar chart illustrating the Log2 surviving fractions (Log2(SF)) of HCC1143 cells transfected with the indicated individual siRNAs and exposed to VE-821 (1 μM) for 5 days. Error bars represent s.d. and P values of <0.001, Student’s t-test, for each siRNA compared with siCon. (h) Western blot illustrating ARID1A protein silencing from experiment (g).
