Figure 2
From: Generating an organ-deficient animal model using a multi-targeted CRISPR-Cas9 system
Induction of DNA DSB in HEK293T by the Cas9-sgRNAms system. (A) DNA DSB assays detected γH2AX accumulation. sgRNAms were driven by the ubiquitously expressing U6-promoter, and Cas9 expression was induced by Doxycycline; the Cas9-sgRNAms system functioned after adding doxycycline to the culture medium. BF means bright field. Blue: nuclei were stained by Hoechst33342, green: Cas9 expression was monitored by EGFP signal, and red: DNA DSBs were indicated by γH2AX accumulation. Images show data from the nt (#2), ms1 (#2), and ms2 (#2) cell lines. The other cell lines showed the same results. The data were quantitatively analyzed as shown in (B). (B) Quantitative analysis of γH2AX intensity. One-way ANOVA, Tukey honestly significant difference test was used in (B). *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. “ns” in (B) means not significant differences.
