Fig. 2: Live cell imaging and mutation analysis with DNA-PKcs inhibition.
From: Improving the sensitivity of in vivo CRISPR off-target detection with DISCOVER-Seq+
a, The proportion of 53BP1 foci relative to BRCA1 as detected by STED in cells exposed to Cas9 targeting a multi-target gRNA with 126 genome-wide target sites. N = 98 cells examined over four independent experiments, P = 0.00018 using two-sided Wilcoxon rank-sum test. b, The number of repair foci (53BP1 or BRCA1) as detected by STED in cells with or without Cas9 (‘+Cas9’ or ‘−Cas9’, respectively), with or without Ku-60648 (‘KU’ versus ‘nD’, respectively), targeting 126 genome-wide sites with a multi-target gRNA. N = 98 cells from four biologically independent replicates, P = 4.44 × 10−10 using two-sided Wilcoxon rank-sum test between ‘−Cas9’ and ‘+Cas9’. Difference in no. of foci in each group was not significant (left, P = 0.15; right, P = 0.95). c,d, Representative images for panel b, (c) with Cas9 or (d) without Cas9. Red labels 53BP1, green labels BRCA1. Scale bar, 5 μm. e, Histogram of indels or no mutations (‘0’) at 48 h after Cas9 editing of ACTB, either without DNA-PKcs inhibitor (‘Cas9, no inhibitor’) or with inhibitor (‘Cas9, Ku-60648’). Untreated cells not exposed to Cas9 shown for reference (‘Untreated (no Cas9)’). Red bar displays mean over two biologically independent replicates. ***P < 0.001. NS, not significant.
