Fig. 6

TaWRKY40 directly binds to the promoter region of TaGAPC1. a Schematic diagram of the probe used for Electrophoretic mobility shift assays (EMSA). Probe1, 2, 3, 4, 5 and 6 contain W-box, whereas in the probe1–1, 2–1, 3–1, 4–1, 5–1 and 6–1, the W-box core sequence was mutated. The mutated bases are indicated in red. b Schematic diagrams of the effector and reporter used for transient transactivation assays in tobacco. c Yeast one-hybrid confirm the interaction. d Transactivation activity reflected by RLUC activity of RLUC/FLUC ratio. Pro35S: GUS was used as an internal control. Quantification was performed by normalizing Firefly luciferase activity to that of Renilla luciferase. e TaWRKY40 binding specific W-box motifs by EMSA. Error bars indicate ±SD (n = 3, from three technical replicates). Significant differences were assessed with one-sided paired t-tests (*, P < 0.05; **, P < 0.01). Three biological experiments were performed, which produced similar results