Fig. 4

SlNAP1 directly binds to the promoters of SlGA2ox1 and SlGA2ox5. A Y1H assays. The CDS of SlNAP1 was fused to the pGADT7 vector, and the promoter fragments of SlGA2ox1 and SlGA2ox5 corresponding to the regions −1500 to −1 were fused to pAbAi vector. B DLR assays. The CDS of SlNAP1 was cloned into the pGreenII 62-SK vector to generate the SlNAP1-62SK effector. The promoters of SlGA2ox1 and SlGA2ox5 (1500-bp upstream of the start codon) were introduced into the pGreenII 0800-LUC vector to generate the ProSlGA2ox1:LUC and ProSlGA2ox5:LUC, reporter constructs, respectively. The constructs were transformed into Agrobacterium tumefaciens strain GV3101. A. tumefaciens was mixed and coinfiltrated into tobacco leaves for transient expression. A DLR assay system was used to measure the ratio of luminescence of firefly LUC to Renilla LUC. Each value represents the means of six biological replicates. *P < 0.05 (Student’s t-test). Y1H yeast one-hybrid, DLR dual-luciferase reporter, CDS coding sequence