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. 2017 Jun 8;18(6):1228. doi: 10.3390/ijms18061228

Figure 3.

Figure 3

(A) Nuclear localization of BrWRKY65 in tobacco leaves. The fusion protein (BrWRKY65-GFP) and positive control were transiently expressed in tobacco leaves respectively via Agrobacterium tumefaciens strain GV3101. GFP signals were captured with a fluorescence microscope after 48 h of injection. Images were photographed in a dark-field for GFP, while the outline of the cell and the merged were taken in a bright field. Bars, 25 μm; (B) Schematics of the reporter and effector constructs. The firefly luciferase (LUC) was drove by the minimal TATA box of the CaMV 35S promoter plus five copies of the GAL4 binding element (5× GAL4), and the CaMV 35S-driving Renilla luciferase (REN) at the same vector was used as an internal control. BrWRKY65 fused with the yeast GAL4 DNA-binding domain (GAL4BD) driven by CaMV 35S, were adopted as the effector; (C) Transcriptional activation activity of BrWRKY65. The trans-activation ability of BrWRKY65 is revealed by the LUC/REN ratio. The ratio of LUC/REN of the empty pBD vector is used as the calibrator (set as 1). At least six independent repeats were assayed for each pair. Compared with the pBD, significant differences at the level of p < 0.05 analyzed by the Student’s t-test, are indicated by different letters above the bar.