Fig. 2

HDR stimulation by NU7441 and KU-0060648 is comparable to other HDR enhancing tools. a Western blot showing knock down efficiency obtained with the siRNAs targeting either Ku70 or Ku80. b Quantitation of genome editing events from cells transfected with pQCiG-TLR and ΔeGFP donor in the presence of 2 μM NU7441, 250 nM KU-0060648, siRNAs targeting Ku70, Ku80, DNA-PKcs or DNA ligase IV, 1 μM Scr7, or a combination of Scr7 and 2 μM NU7441 or 250 nM KU-0060648. The HDR and NHEJ values are relative to those obtained with Cas9, sgRNA, and ΔeGFP donor in the presence of vehicle (DMSO). Results are from biological replicates performed in technical duplicates. Significance (relative to vehicle) was calculated using the Student’s t-test: *p ≤0.05; **p ≤0.01; ns, not significant. c Western blot showing expression of Adenovirus 5 proteins E1B55K and E4orf6 following transfection into the 293/TLR line. d Quantitation of genome editing efficiencies was as in B except that adenovirus 5 proteins E1B55K and E4orf6 expression vectors were co-transfected with pQCiG-TLR and ΔeGFP donor plasmids and cultured in presence or absence of 2 μM NU7441 or 250 nM KU-0060648. N = 4; error bars represent S.D. Results are from biological replicates performed in technical duplicates. Significance (relative to vehicle) was calculated using the Student’s t-test: *p ≤0.05; **p ≤0.01; ns, not significant