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. 2017 Feb 17;8:14432. doi: 10.1038/ncomms14432

Figure 1. BRCA2 deficient cells are highly sensitive to CX-5461 in different human and murine cell types.

Figure 1

(a) The colony formation capacity of BRCA2−/− HCT116 cells was greatly reduced by treatment with CX-5461. Experiments were repeated twice with similar results. Scale bar, 1 cm. (b) The hypersensitivity of BRCA2−/− cells to CX-5461 in HCT116 validated by WST-1 assay. Representative experiment #3 (see Supplementary Fig. 1a for full experimental panels, n=9) is displayed as individual data points and as fitted sigmoid dose response curves (green for BRCA2 deficient and red for BRCA2 wild type). Dashed vertical lines are the IC50. (c) CX-5461 induced more apoptosis in BRCA2−/− cells as indicated by FACS analysis. A representative result is shown on the left panel and the right panel shows mean apoptotic fraction with 95% CI of cells in early apoptosis under different drug concentrations. ***P<0.0001, t-test; n=3, 2 or more replicates per condition. See Supplementary Table 5 for more statistical analyses. (d) BRCA2−/− in DLD1 isogenic cell line pairs displayed hypersensitivity to CX-5461 by WST-1 assay. Representative experiment #3 is shown (see Supplementary Fig. 2a for full experimental panels; n=4). Green fitted sigmoid curves are for BRCA2 homozygous (HOM) and red for BRCA2 wild type and heterozygous (HET). (e) BRCA2 deficient ovarian cancer PEO1 cells exhibited increased sensitivity to CX-5461 relative to BRCA2 proficient C4-2 cells by WST-1 assay. A representative experiment #1 is displayed (see Supplementary Fig. 2b for full experimental panels, n=3). (f) RNAi knockdown of BRCA2 increased sensitivity to CX-5461 in p53−/− HCT116 cells by WST-1 assay (4 days in drug). The results of all three experiments are summarized by the green (BRCA2 knockdown) and red (non-targeting control) super-smoother fit lines. (g) 45S pre-rRNA level measured by RT-PCR after CX-5461, CX-3543 and BMH-21 treatment in WT and BRCA2−/− HCT116 cells. Drug incubation time was 24 h. Fold change estimates and unadjusted 95% CIs of 45s pre-rRNA levels under drug treatment condition versus vehicle control are shown. P values (by F-test) are shown in Supplementary Table 7. (h) BRCA2 knockout cells are not more sensitive to BMH-21 in HCT116 through WST-1 assay. One representative experimental result is shown (more replicates are shown in Supplementary Fig. 3a).