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. 2014 Oct 27;33(23):2860–2879. doi: 10.15252/embj.201489017

Figure 2. Cdh1 depletion sensitizes to DNA-damaging agents and affects recruitment of DNA repair components.

Figure 2

  1. MCF7 cells were infected with the indicated shRNAs and selected with puromycin. pLL-GFP- or pLL-Cdh1-infected MCF7 cells were plated in 6-well plates and subsequently irradiated with the indicated amounts of ionizing radiation. Surviving colonies were stained, and relative amounts of colony numbers compared to non-irradiated cells are shown. Averages and standard deviations of three independent experiments are shown.
  2. pLL-GFP- or pLL-Cdh1-infected MCF7 cells were plated in 96-well plates and subsequently treated with the indicated amounts of doxorubicine for 4 days. Cellular viability was assessed using MTT conversion, and untreated cells were used as a reference. Averages and standard deviations of three independent experiments are shown.
  3. At 48 h after siRNA transfection, U2OS-FUCCI cells were irradiated (2 Gy). At 2 or 5 h after treatment, whole-cell extracts were prepared and analyzed by Western blotting with the indicated antibodies.
  4. U2OS-FUCCI cells treated as in (C) were prepared for 53BP1 and Rad51 immunofluorescence. Graphs show the amounts of 53BP1 or Rad51 foci in S/G2 cells. At least 120 cells from three independent experiments were counted for each condition, and data are presented as box plots with whiskers representing the minimal and maximal values. Unpaired Student's t-tests (two-sided) were done to compare control-depleted and Cdh1-depleted conditions. Representative images can be found in Supplementary Fig S2B and C.