Genomewide identification of ANAC060 binding targets

In this study, genome-wide binding analysis of ANAC060 was undertaken through chromatin immunoprecipitation sequencing assays using the transgenic lines expressing functional ANAC060-GFP fusion protein in anac060 background. A total of 3282 genes were identified to associate with ANAC060 binding sites. The G-box binding motif was highly enriched in the ANAC060 binding peaks. Two grams of 10-d-old whole seedlings of 35S:ANAC060-GFP/anac060 transgenic line were used for ChIP assays.The wild type (Col) grown under the same conditions were used as the control. Polyclonal anti-GFP antibodies (Abcam, ab290) were used to immunoprecipitate the chromatin complexes. Sequencing libraries were constructed according to the Illumina’s instructions.