Supplementary Figure 4: Control immunoblots and assessment of AAV-i53 vectors (related to Fig. 4) | Nature Biotechnology

Supplementary Figure 4: Control immunoblots and assessment of AAV-i53 vectors (related to Fig. 4)

From: Inhibition of 53BP1 favors homology-dependent DNA repair and increases CRISPR–Cas9 genome-editing efficiency

Supplementary Figure 4

a,Immunoblots of whole cell lysates prepared from U2OS DR-GFP cells transfected with the indicated siRNAs along with an I-SceI expression vector and probed with the indicated antibodies. This blot relates to the experiment shown in Fig. 4b. b, Immunoblots of whole cell lysates prepared from U2OS DR-GFP cells transfected with vectors expressing Flag-tagged i53, its DM mutant or an empty vector along with an I-SceI expression vector and probed with the indicated antibodies. This blot relates to the experiment shown in Fig. 4c. c, Control immunoblot of the experiment shown in Fig. 4d. d,e U2OS DR-GFP cells transduced with AAV particles coding for Flag-i53 (i53) or Flag-i53-DM (i53-DM) and then subjected to the DR-GFP assay following transfection of the I-SceI expression vector. In (d) the percentage of GFP-positive cells was determined 48 h post- transfection for each condition. Each pair of points are matched biological replicates; N=3. Control immunoblots showing i53 and i53-DM expression are shown in (e). Please note that the blots shown are cropped and the uncropped versions can be found as Supplementary Information.

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