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. 2007 May 5;35(11):3535–3550. doi: 10.1093/nar/gkm195

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© 2007 The Author(s)

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — Short- and long-term gene silencing for MRN components. (A and B) 45, 69 and 76 days after transfection, Rad50^KD HeLa clones were analyzed by immunocytochemical staining (magnification ×315) or western blot. 1: Control (pBD650; day 189); 2: XPC^KD cl. 21 (pBD634; day 376); 3: XPA^KD clone 6 (pBD695; day 436); 4: XRCC4^KD clone 9 (pBD694; day 107); 5: Control (pBD650; day 31); 6: Rad50^KD clone 1 (pBD872; day 76); 7: Rad50^KD clone 1 (day 45); 8: Rad50^KD clone 2 (day 45); 9: Rad50^KD clone 3 (day 45); 10: Rad50^KD clone 4 (day 45); 11: Rad50^KD clone 5 (day 45). (C) Transient gene silencing: 6 days after transfection, MRE11^KD (two pEBVsiRNA plasmids), Rad50^KD (one plasmid) and NBS1^KD (two plasmids) cell populations were analyzed by western blot. 1: Control (pBD650; day 6); 2: MRE11^KD (pBD930; day 6); 3: MRE11^KD (pBD931; day 6); 4: NBS1^KD (pBD932; day 6); 5: NBS1^KD (pBD934; day 6); 6: ATM^KD (pBD935; day 6); 7: ATM^KD (pBD936; day 6); 8: ATM^KD (pBD937; day 6); 9: PARG^KD (pBD812; day 6); 10: Rad50^KD (pBD872; day 6); 11: Control (pBD650; day 6); 12: Rad50^KD (day 120). (D) Long-term gene silencing: Clones were picked up from cell populations described in the panel ‘C’ and propagated for several months in culture. 1: Control (pBD650; day 238); 2: MRE11^KD cell population (pBD930; day 33); 3: MRE11^KD clone 32 (pBD930; day 132); 4: Rad50^KD clone 1 (pBD872; day 149); 5: NBS1^KD clone 11 (pBD932; day 172); 6: DNA PKcs^KD clone 1 (pBD743; day 391); 7: Ligase III^KD clone 11 (pBD941; day 180).