Mutagenesis profiles revealed by 454 sequencing. (A) The total mutation (insertion, deletion, and nucleotide substitution) frequency for each sample was scored. (B) Proportion of deletions in total mutation events in different genotypes under different treatments. (C) Proportion of insertions in total mutation events in different genotypes under different treatments. (D) Relative ratio of deletion events to insertion events in different genotypes under different treatments. Protoplasts derived from Col, ku70, lig4, and smc6b plants and carrying the same ADH1-ZFN-4 transgene were treated with donor, estradiol, or donor and estradiol. Two (donor treatment) or three (estradiol, donor and estradiol) independent biological replicates were subjected to 454 high-throughput sequencing and analysis. (Error bars) Standard errors. In all panels, statistically significant differences between the mutants and Col (WT) are indicated by asterisks (P < 0.05, t-test).