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. 2017 Aug 22;13(8):e1006955. doi: 10.1371/journal.pgen.1006955

Fig 9. OsEIL1 directly binds to YUC8/REIN7 promoter region.

Fig 9

(A) Schematic diagrams of putative EIN3 binding site (EBS) in the promoter of YCU8/REIN7. Black boxes indicate the positions of the EBS. P1-P4 are fragments of the YUC8/REIN7 promoter. (B) Anti-myc ChIP assays with DNA from 3-d-old etiolated seedling roots of Nip and overexpressing OsEIL1 with myc-tag (EIL1-myc) transgenic plants. (C) EMSA assay for binding to EBS sequence in the promoter of YUC8/REIN7 by OsEIL1 protein in vitro. Glutathione S-transferase (GST)-tagged OsEIL1 N-terminal fusion protein was incubated with biotin-labeled DNA fragments (Probe). Competition for the biotin-labeled promoter region was done by adding an excess of unlabeled probe (Competitor). Three biological replicates were performed with similar results. (D) The activation of OsEIL1 on the promoter activity of YUC8/REIN7 by transient expression assay in tobacco leaves. ‘EV’ represents empty vector. Three biological replicates were performed with similar results. (E) Quantitative analysis of luminescence intensity for each comparison in (D).