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. 2014 Aug 6;42(15):9771–9780. doi: 10.1093/nar/gku650

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© The Author(s) 2014. 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 5. — Delayed resolution of repair foci. Vectors encoding control or PSPC1 miRNA were introduced by electroporation into wild-type (+/0) or NONO-deficient (gt/0) MEFs. Cells were mock irradiated or exposed to 1 Gy of ¹³⁷Cs γ-rays. At indicated times following irradiation, cells were fixed and analyzed by indirect immunofluorescence using anti-γ-H2AX primary and red secondary antibody and DAPI counterstain. (A, C, E) Representative fields showing γ-H2AX/DAPI channels (top row) or γ-H2AX/EmGFP/DAPI-merged images in NONO-deficient (gt/0) MEFs. Images were collected for non-irradiated control cells or for cells that were irradiated and allowed to recover for indicated times. Cell morphology and foci appearance were indistinguishable in wild-type (+/0) and NONO-deficient (gt/0) MEFs; only the latter are shown in the figure. Scale bar denotes 10 μm. (B, D, F) Foci per cell in populations corresponding to panels (A, C, E). A total of 30 nuclei were scored per experimental group. Blue symbols depict score for individual cells; red bar depicts mean. Treatment of NONO-deficient cells with PSPC1 RNA resulted in a significant increase in residual foci at 4 h post-irradiation (P < 0.01).