Figure 3.

Generation of MCM10 mutant cell lines on a HeLa background. (A) Schematic showing the strategy used to construct an AID mutant using CRISPR-KO and bicistronic pAID5 plasmids. The CRISPR-KO plasmid expresses a Cas9–gRNA complex for gene knockout. The bicistronic rescue plasmid is integrated into the genome by the action of TOL2 transposase and expresses both OsTIR1 and mAID-fused protein. (B) Immunoblot showing endogenous MCM10 and mAID–MCM10. HeLa clones were treated with 500 μM IAA for 2 h and protein extracts were separated by SDS-PAGE. Tubulin is a loading control.