Fig. 3.

PIFs directly bind to MIR156 promoters. a The transcript levels of mature MIR156 decline in WT in response to EOD-FR treatment. Eight-day-old (left panel) or 3-week-old (right panel) WT plants grown under WL were treated with or without EOD-FR for 6 days before harvested for RNA extraction. Values given are mean ± SD (n = 3). *P < 0.05 and **P < 0.01 by the Student’s t-test. b Y1H assay showing direct binding of PIFs to the PBE-box or G-box motifs in several MIR156 promoters. The length and PIF-binding elements of each promoter fragment is shown in Supplementary Fig. 6. c EMSA showing that the GST-PIF5 bHLH domain recombinant protein binds to biotin-labeled probes of MIR156E (up panel) and MIR156F (down panel). d ChIP assay of MIR156 in 2-week-old 35S::PIF5-HA seedlings grown under high R:FR conditions and treated with EOD-FR for one time before harvesting. Values given are mean ± SD (n = 3). **P < 0.01 by the Student’s t-test