Figure 4: Biological penetrance of mitotic perturbation determines the timing and magnitude of mitosis-born DNA breakage.
From: Profiling DNA damage response following mitotic perturbations
(a) A representative image (n=50) of U-2-OS cells treated with siRNA to CDC27 and immunostained for γ-H2AX. Nuclear DNA was counterstained with DAPI. Scale bar, 10 μm. The bar charts (left) show γ-H2AX-decorated DSBs in mitotic cells (n=250) in the indicated times after siRNA transfection. (b) U-2-OS cells were treated with siRNA to PLK1 and analysed as in a. Scale bar, 10 μm. (c) Representative image galleries (n=25) of U-2-OS cells expressing mRFP-53BP1, treated with the indicated siRNAs, and analysed by time-lapse microscopy. Average mitotic duration (n=25) for each siRNA treatment is indicated. Numbers indicate time (h) after siRNA transfection. Scale bars, 10 μm. (d) Representative image galleries (n=25) of U-2-OS cells expressing mRFP-53BP1, treated with the indicated siRNAs, and analysed as in c. Average mitotic duration (n=25) for each siRNA treatment is indicated. Numbers indicate time (h) after siRNA transfection. Scale bars, 10 μm.
