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Fig. 4 | BMC Plant Biology

Fig. 4

From: LATE ELONGATED HYPOCOTYL regulates photoperiodic flowering via the circadian clock in Arabidopsis

Fig. 4

LHY does not bind to FT promoter. a Genomic structure of FT gene. (Left panel) Gray box indicates the gene promoter region. Black boxes indicate exons, and white boxes indicate untranslated regions. CBS, CCA1-binding sequence. EE, evening element. NB, non-binding sequence. (Right panel) CBS and EE sequences are listed. b ChIP assays on binding of CCA1 and LHY to FT promoter. A MYC-coding sequence was fused in-frame to the 5′ end of the CCA1-coding sequence and the 3′ end of the LHY-coding sequence, and the gene fusions were overexpressed driven by the Cauliflower Mosaic Virus (CaMV) 35S promoter in Col-0 plants, resulting in 35S:MYC-CCA1 and 35S:LHY-MYC, respectively. Chromatins were prepared from 7-day-old whole plants grown on MS-agar plates and immunoprecipitated using an anti-MYC antibody. Fragmented genomic DNA was eluted from the protein-DNA complexes and subjected to quantitative PCR. Biological triplicates were averaged and statistically treated using Student t-test (*P < 0.01). Bars indicate SE. The promoter sequences of eIF4a and TUB genes were included as negative controls in the assays. The promoter sequence of TIMING OF CAB EXPRESSION 1 (TOC1) gene containing EE was included as positive control [35]. c Suppression of FT transcription by CCA1. The reporter and effector constructs are illustrated (left panel). Transient GUS expression assays were performed using Arabidopsis protoplasts (right panel). Five measurements were averaged and statistically treated (t-test, *P < 0.01). Bars indicate SE

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