Fig. 5. SOC1 directly regulates AGL24. (A,B) Relative temporal expression of AGL24 (A) and AP1 (B) in developing Arabidopsis seedlings of different genetic background under long-day conditions. (C) Relative temporal expression of AGL24 in the aerial part without leaf and leaf of soc1-2 and wild-type seedlings. Transcript levels in A-C were determined by quantitative real-time PCR analyses of three independently collected samples. Results were normalized against the expression of TUB2. Error bars indicate s.d. (D) Schematic of the AGL24 genomic region. Arrowheads indicate the sites containing either single mismatch or perfect match with the consensus binding sequence (CArG box) of MADS-domain proteins. Four PCR fragments corresponding to the DNA sequences near these CArG boxes were designed for ChIP analysis. (E) ChIP enrichment test shows the binding of SOC1-9myc to the region near the number 1 fragment indicated in D. (F) Schematic of the Pro_AGL24:GUS construct. Two native CArG boxes within the number 1 fragment identified in D and E were mutated as indicated. (G) Representative GUS staining in 12-day-old transformants containing Pro_AGL24:GUS and its derived constructs with the mutated CArG boxes (M-2003 and M-2039). (H) Distribution of relative GUS staining intensity in the transformants containing M-2003 and M-2039. (I) GUS staining of Pro_AGL24:GUS and M-2039 in the wild-type (left) and 35S:SOC1 (right) background. Representative lines of transformants containing Pro_AGL24:GUS and M-2039 were crossed with 35S:SOC1, and GUS staining of 4-day-old F1 plants is shown on the right. (J) Distribution of flowering time in T1 transgenic plants carrying the wild-type AGL24 gene and its mutated forms (M-2003 and M-2039) in the agl24-1 mutant background.