Figure 2: DNA damage is induced in cells with CX-5461 and CX-3543 treatment.
From: CX-5461 is a DNA G-quadruplex stabilizer with selective lethality in BRCA1/2 deficient tumours
(a) The formation of γ-H2AX, 53BP1, RPA and RAD51 foci was monitored upon CX-5461, CX-3543 and BMH-21 treatment at 10−7 M in U2OS cells. Drug treatment time is 24 h for all drugs. Scale bar, 10 μM. (b) Beanplots of U2OS cells showing five or more γ-H2AX foci for the indicated drug treatment condition after 24 h. **P<0.01 (one tailed randomization tests adjusted for multiple comparisons relative to vehicle control); n=2 experiments, (35–150 cells were counted each time per each condition). DMSO was the vehicle control solvent for CX-3543 and NaH2PO4 was the vehicle control solvent for BMH-21 and CX-5461. (c) Beanplots of HCT116 cells showing three or more 53BP1 foci for the indicated drug treatment conditions after 24 h. **P<0.01, ***P<0.0001 (one-tailed randomization tests adjusted for multiple comparisons relative to vehicle control); n=3 experiments; >100 cells per replicate condition. (d) Quantification of alkaline comet and neutral comet assay result upon CX-5461 treatment (30 min) in HCT116. Tail moments were determined as described in Methods; (n=3 experiments, 100 cells were counted in each replica), mean and 95% CI are shown. Right panels show pair-wise comparison of condition contrasts and 95% CIs of tail moment difference. (e) Representative images of alkaline comet assay of cells treated with CX-5461 (10−6 M, 30 min) and no drug control. Scale bar, 20 μM.
