Aplf knockdown does not compromise DNA repair competency in iPSCs.

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Aplf knockdown does not compromise DNA repair competency in iPSCs. Aplf knockdown does not compromise DNA repair competency in iPSCs. (A) Control and Aplf-kd MEFs at day 9 of reprogramming were exposed to different concentrations of actinomycin D for 16 h. Annexin-V-positive cells were counted to determine the early apoptotic cell population. (B) The same set of cells described in A were treated with 10 µM etoposide for 4 h to induce DNA damage. After 4 h, the etoposide-containing medium was replaced with normal ESC culture medium, and cells were allowed to recover. At 0 h and at different time points of recovery, cells were fixed and analyzed for the presence of γH2AX foci by immunofluorescence staining. Nuclei with ≥5 foci were counted as positive. NT, non-treated. (C) C21 iPSCs and E14 ESCs at passage 26 were grown in a 24-well plate for 16 h and treated with 10 µM etoposide in a similar manner to that described above. At 0 h and at different points of recovery, cells were fixed and analyzed for γH2AX levels by immunofluorescence. Representative colonies are shown. Nucleus, Hoechst 33258, blue. Error bars are s.e.m., n=3, *P<0.05 (Student's t-test). Khaja Mohieddin Syed et al. J Cell Sci 2016;129:4576-4591 © 2016. Published by The Company of Biologists Ltd