Figure 5.

Interaction of OsASR2 with the GT‐1 cis‐element. (a) Interaction of OsASR2 with the GT‐1 cis‐element by the yeast selection system. Y1HGold yeast strains carrying empty vector pGADT7 or pGADT7‐OsASR2 were grown overnight at 30 °C. Cultures were diluted to OD ₆₀₀ = 1.0, and then, serial 10‐fold dilutions were spotted onto medium lacking Ura and Leu in the presence of 150 ng/mL AbA. (b) EMSA using the recombinant OsASR2 protein and the GT‐1 cis‐element. Lane 1 contained only the free probe. Lane 2 contained the purified OsASR2 protein and the probe. For the competitive EMSA, the purified OsASR2 protein was pre‐incubated with 5 (lane 3)‐, 25 (lane 4)‐ or 50 (lane 5)‐fold molar excess of unlabelled GT‐1 cis‐element before the addition of the biotin‐labelled GT‐1 cis‐element. For the mutant EMSA, the purified OsASR2 protein was incubated with the labelled mutant GT‐1 cis‐element. (c) The positions of primers used in the ChIP‐qPCR experiment. (d) Binding of OsASR2 to the GT‐1 cis‐element in a ChIP‐qPCR assay. (e) The GT‐1 binding motif identified in OsASR2 binding peaks in ChIP‐seq assay. (f) OsASR2 biding profile in the promoter of Os2H16. The open arrowhead refers to the GT‐1 around the peak summit. The red vertical line denotes the peak summit.