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Figure 6


Fig. 6. Silencing of NBS1 or MDC1 increases rAAV transduction. (A) β-galactosidase activity measured as relative light units (RLU) in lysates from MRC5 cells after treatment with siRNAs against NBS1 (siNbs1) or luciferase (siLuc), followed by transduction with AAV-LacZ at three multiplicities of infection [m.o.i.; 10000, 5000 and 2500 viral genome particles (vgp) per cell]. Cells were either not treated or were treated overnight with 5 mM HU. (B) Same as in A but using HeLa cells. (C) Efficiency of AAV-LacZ (10,000 vgp/cell) transduction (measured as above) of NBS1-ILB1 cells (mutated in both copies of the NBS1 gene) and NBS1-ILB1/NBS1 cells (in which NBS1 function had been complemented by transfection of full length NBS1). (D) Same as in panel A but using AT5 cells, without HU treatment. (E) Flow cytometry analysis of retrovirally transduced HeLa cells expressing shRNAs against MDC1 (pSR-Mdc1) or β-gal (pSR-LacZ) after infection with AAV-GFP. The upper panel shows the levels of MDC1 protein in the two cell lines. (F) Flow cytometry analysis of HeLa cells transfected with siRNAs against MDC1 (siMdc1) or luciferase. The upper panel shows the levels of MDC1 protein 60 hours after siRNA transfection (24 hours after AAV-GFP transduction).





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