Research Paper Volume 3, Issue 4 pp 395—406

Figure 3. Depletion of Ku70/80 in CCL75.1 cells does not affect telomere length nor overhangs signal. (A) Terminal Restriction Fragment (TRF) analysis. CCL75.1 cells transduced with lentiviruses for the conditional expression of shRNAs targeting Ku70 and Ku80 or GFP were harvested at 7 (shKu70/80) and 10 days (shKu70/80 and shGFP) after induction with DOX. Equal amounts of genomic DNA digested with HinfI and RsaI were separated by electrophoresis on a 0.8% agarose gel and analyzed by Southern blotting with a radiolabeled (CCCTAA)₄probe. The molecular mass standards shown on the right lane were generated by digestion of lambda DNA with the restriction endonuclease HindIII. (B) Telomere 3′ overhang signal analysis. CCL75.1 cells transduced with lentiviruses for the conditional expression of shRNAs targeting either Ku70/80 or GFP were harvested at 7 (shKu70/80) and 10 days (shKu70/80 and shGFP) after induction with DOX. Equal amounts of genomic DNA digested with HinfI and RsaI were separated by electrophoresis on a 0.8% agarose gel. The gel was dried under native conditions and then hybridized to a radiolabeled (CCCTAA)₄probe.