Fig. 5

CmNAC25 activates the promoters of CmMYB6 and CmDFR. A, B Interactions between CmNAC25 and different fragments of CmMYB6 or CmDFR promoter in Y1H assays. Yeast cells co-transformed with pGADT7-Gus and pHIS2 constructs were used as negative controls. SD/-L/T/H indicates Leu, Trp, and His synthetic dropout medium. C EMSA assay showing that CmNAC25 directly binds to CmMYB6 promoter at the ACGT element located at − 89 ~  − 85 bp. * means non-specific binding band. D, E Dual-luciferase assays showing that CmNAC25 activates the promoter activities of CmMYB6 and CmDFR. LUC driven by the CmMYB6 or CmDFR promoter was used as a reporter. The pORE-R4-35S::NAC25 construct was an effector, and the empty vector (pORE-R4) served as a control. The LUC/REN ratio of controls was set as 1. Error bars indicate the SD of three biological replicates. Samples denoted by asterisks indicate significant differences (p < 0.05, t-test)