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. 2019 Nov 20;21(12):1143–1150. doi: 10.1016/j.neo.2019.09.002

Figure 1.

Figure 1

Decreased DOX-induced DNA damage in APC-deficient cells. A) Protein levels of phosphorylated H2AX (yH2AX), a marker of DNA damage, was measured in MMTV-PyMT;ApcMin/+ and MMTV-PyMT;Apc+/+ cells following 24 hr treatment of chemotherapeutic agents, cisplatin (CIS), doxorubicin (DOX), and paclitaxel (PTX). B) Quantification of western blots show that yH2AX was induced after DOX and PTX treatment in MMTV-PyMT;Apc+/+ cells, but not in MMTV-PyMT;ApcMin/+ cells. In contrast CIS treatment induced equal damage in both cell lines. C) Representative images of yH2AX immunofluorescence (IF) in DOX and PTX-treated MMTV-PyMT;Apc+/+ and MMTV-PyMT;ApcMin/+ cells. White arrows depict yH2AX positive cells, and the scale bar is 20 microns. D) Quantification of yH2AX IF images show an increased percentage of yH2AX in DOX-treated MMTV-PyMT;Apc+/+ cells, which is reduced in DOX treated MMTV-PyMT;ApcMin/+ cells. Each experiment was repeated 3 times, at least 150 cells were counted per condition in each experiment. E) MMTV-PyMT;Apc+/+ and MMTV-PyMT;ApcMin/+ cells were treated with DOX or PTX for 24 hours and DNA damage was assessed by the comet assay. The tail moment was measured and showed that DOX-treated MMTV-PyMT;Apc+/+ cells have increased tail moment compared to solvent control. DOX-treated MMTV-PyMT;ApcMin/+ cells show no change compared to solvent control, and were significantly decreased compared to the DOX-treated MMTV-PyMT;Apc+/+ cells. The data are shown as means ± SD; *P < 0.05 comparing MMTV-PyMT;ApcMin/+ or MMTV-PyMT;Apc+/+ cells treated to solvent control and **P < 0.05 comparing MMTV-PyMT;ApcMin/+ to MMTV-PyMT;Apc+/+ cells.