Fig. 4

Two regions of the EAS4 promoter are required and sufficient for activation by wounding: Agrobacterium cells carrying GUS driven by the respective promoters were mixed with those carrying LUC driven by 35S promoter, and infiltrated into N. benthamiana leaves. At 40–48 h after infiltration, the leaves were wounded, and harvested at the times indicated after wounding. Transcript levels of GUS, LUC, and Nbactin2 were quantified by RT-qPCR, and the level of GUS was doubly normalized to the levels of Nbactin2 and LUC as internal and infection standards, respectively. a Shown is a schematic representation of the EAS4 promoter. b 5′-Deletion analysis of the EAS4 promoter. Values are means with standard deviations of six to nine biological replicates. c Deletion analysis of 160p. Values are means with standard deviations of three to six biological replicates. d Gain-of-function analysis of the EAS4 promoter. Four tandem repeats of respective regions of the EAS4 promoter were fused to a 35S minimal promoter. Values are means with standard deviations of three biological replicates. Significant differences between 0 h and 12 h were determined by Student’s t-test using Excel 2013 software (**P < 0.01, *P < 0.05)