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. 2015 Sep 8;44(1):117–133. doi: 10.1093/nar/gkv885

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© The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 4. — EPC1 inhibits DNA damage-induced apoptosis and promotes cell motility. (A) Percentage apoptotic SK-Mel-147 (left) and UMUC3 (right) cells treated with cisplatin (cDDP, 30 μM) for 48 h upon EPC1 knockdown. Cells were infected with lentivirus expressing sh.EPC1 or scrambled RNA (sh.control) and then treated with cDDP. Apoptosis was measured by PI staining and analyzed by FACS. (B) Percentage cell death of Saos-2 (left) and SK-Mel-29 (right) cells treated with cDDP for 48 h after overexpression of EPC1. Western blot confirmed EPC1 expression. (C) Saos2.ER-E2F1 cells were treated with 4OHT or solvent and apoptosis was measured with and without ablation of EPC1. (D) E2F1 or scrambled shRNA were expressed in SK-Mel-147 cells. Simultaneously, EPC1 was overexpressed by Ad.EPC1 infection for 24 h. Cells were treated with cDDP for 36 h and analyzed by FACS. (E) SK-Mel-147 cells were infected with lentivirus expressing scrambled or EPC1 shRNA for 96 h and analyzed in a scratch migration assay. Representative phase contrast microscopy images of the migration assays at 0, 24 and 48 h after gap creation are shown (left). Quantitative data are shown (right). (F) E2F1 or scrambled shRNA was expressed in metastatic bladder cancer cells T24 (upper) and UMUC3 (lower) followed by EPC1 overexpression by Ad.EPC1 infection. The migratory potential of tumor cells was analyzed by cell migration assays. Representative images at 0 and 24 h after gap creation are shown. Quantitative data are presented as bar graphs. Bars represent the mean ± SD of three independent experiments. Statistical significance was determined by two-sided Student's t-test. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001.