Figure 3. ATM-mutant p53 axis of the DNA damage response (DDR) promotes cell migration and invasion in MDA-MB-231 cells.

(A) ATM or mutant p53 depletion, as well as co-depletion, impairs cell motility similarly. Wound-healing assays were performed with the indicated siRNAs as in Figure 2A. (B) siRNA depletions and quantification of wound healing for (A). (C) Real-time analysis of cell dynamics in siATM, simutant-p53 and co-depleted cells. Experiments performed as in Figure 1G with indicated siRNAs. Right: ATM and mutant p53 levels in cell samples. (D) Live cell imaging of cell migration defects in ATM and mutant p53 depleted MDA-MB-231 cells. Experiments were performed and analyzed as in Figure 2F. Scale bar, 20 μm. (E) Quantification of individual cell speed (μm/min) and cell path (μm) from (D). Cell parameters were quantified as in Figure 2G. Error bars = SD. *** p-value <0.0001, unpaired two-tailed t-test.

DOI: http://dx.doi.org/10.7554/eLife.07270.009

Figure 3—figure supplement 1. Analysis of ATM and mutant p53 functions in human breast cancer cells.

(A) ATM depletion impairs cell migration. Cells transfected with indicated siRNA were analyzed by wound-healing assays to determine cell motility. Right: Quantification of wound healing and ATM siRNA knockdown efficiency. (B) Reduction of ATM and/or p53 reduces cell migration. Cells were treated and analyzed with the indicated siRNAs as Figure 3A. Quantification of wound healing and depletion analysis is in right. (C) Treatment of MDA-MB-231 cells with ATM kinase inhibitor reduces mutant p53-pS35 compared to untreated cells. Experiments were performed as in Figure 1F.

DOI: http://dx.doi.org/10.7554/eLife.07270.010