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. 2020 Sep 21;9(9):1246. doi: 10.3390/plants9091246

Figure 5.

Figure 5

PIF4 directly binds to ERF1 promoter and suppresses its expression. (a) Schematic diagram of the ERF1 promoter. The solid line indicates the position of G-box in the promoter. (b) ChIP-qPCR, using pPIF4-PIF4-MYC/pifq plants, shows that PIF4 binds to the ERF1 promoter region containing the G-box in vivo. The error bars indicate the SD of three independent experiments. Asterisks indicate Student’s t-test significant differences (* p < 0.05). (c) EMSA assay shows that GST-PIF4 bHLH recombinant protein binds to the promoter of ERF1 in vitro. DNA fragments of ERF1 promoter were synthesized with normal G-box motif (G-box probe) and mutated G-box motif (mutant probe), and labeled with biotin. The GST protein was used as negative control. (d) The diagram of the reporter and effectors used in the protoplast transfection assays. Transient expression assays indicate that PIFs repress ERF1 expression. The error bars indicate SD of three independent experiments. The different letters above columns indicate significant differences (one-way ANOVA; p < 0.05).