Figure 6: The FRS7–FRS12 complex binds and represses genes responsible for diurnal growth and flowering.

(a) Venn diagram comparing the FRS12 physically bound genes to the transcriptionally regulated genes in the Pro35S:FRS12-GR-1 line. (b) Representations of the TChAP-Seq FRS12 binding peaks located at the PIF4, PIL1 and GI promoters. The reads are piled up in forward reads above the axis in green and reverse reads below the axis in blue. Total coverage is indicated in yellow. Coloured bars on top of promoters designate protein–DNA-binding motifs. Black lines below promoters highlight selected regions for ChIP-qPCR analysis. (c) Logo representation of the FRS12 DNA-binding elements FRB1, FRB2 and FRB3, identified by de novo motif enrichment analysis. (d) ChIP-qPCR assay of selected fragments in the PIF4, PIL1 and GI promoters. A transgenic Arabidopsis line expressing ProFRS12:FRS12-HA was grown for 10 days in SD conditions and harvested at ZT8 for analysis. Enrichment values were normalized to respective inputs and represented relative to Col-0 wt plants (background control). Values represent the mean of three biological replicates±s.e.m.; *P<0.05, ***P<0.001, t test. (e–g) Transient expression assays in N. benthamiana showing the trans-repression of the PIF4, PIL1 and GI promoters by FRS7 and FRS12. Values represent the mean of eight technical replicates±s.e.m.; *P<0.05, ***P<0.001, t test.