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Figure 3


Fig. 3. Colocalization of rAAV foci with DNA-damage-response proteins. (A-E) MRC/GFP-LacR or HeLa/GFP-LacR (where indicated) cells were transduced with AAVLacO.14 and, after 24 hours, fixed and immunostained with anti-NBS1 (Nbs1; A), P-S343-NBS1 (B), MRE11 (Mre11; C), RAD50 (Rad50; D) and MDC1 (Mdc1; E) antibodies. HU and campto: cells that were treated with hydroxyurea (1 mM overnight) or camptothecin (1 nM for 6 hours) prior to AAVLacO.14 transduction. The dotted rectangles indicate the portions of the merged figures enlarged in the rightmost panels. Representative cells of at least 50 analyzed per condition are shown. (F) Number of rAAV foci colocalizing with MRN and MDC1 foci. Quantifications were performed for cells treated with HU (NBS1, P-S343-NBS1, MRE11 and RAD50) or camptothecin (MDC1) as in A-E.