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Figure 4 | BMC Pharmacology

Figure 4

From: Enhanced sensitivity to cisplatin and gemcitabine in Brca1-deficient murine mammary epithelial cells

Figure 4

Effect of Brca1 -deficiency on NER and Double-strand DNA Break Repair. In (a), GGR of CPDs in Brca1+/+ (black circle) and Brca1-/- (white square) cells was measured using an immunoslot blot assay. Cells were exposed to 10 J/m2 UV-irradiation and collected at the indicated times. DNA repair was expressed as a percentage relative to control. Data from triplicate DNA samples from three different biological experiments were expressed as an average ± S.E.M. In (b), sensitivity to UV-irradiation was determined by MTT assay for Brca1+/+ (black circle) and Brca1-/- (white square) cells. In (c), damage-induced expression of Xpc mRNA, an NER gene involved in DNA damage recognition, in Brca1+/+ and Brca1-/- cells was measured using RT-qPCR. Brca1+/+ and Brca1-/- cells were exposed to 10 J/m2 of UV and either harvested immediately (control) or incubated in media and harvested 24 h later. In (d), expression of Xpc mRNA following 24 hours of treatment with 0.1 μM cisplatin or 0.01 μM gemcitabine in Brca1+/+ and Brca1-/- cells was measured using RT-qPCR. Data were calculated relative to the untreated control and expressed as the average of three experiments ± S.E.M. In (e), DNA strand breaks were measured at 24 hours following treatment with 0.1 μM cisplatin, 0.01 μM gemcitabine, or 0.1 μM doxorubicin in Brca1+/+ and Brca1-/- cells using the alkaline comet assay. Comet tails indicate DNA damage. Unless indicated otherwise, data were expressed as an average of triplicate readings ± S.D. **, p < 0.01; *, p < 0.05.

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