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. 2019 Mar 8;180(1):621–633. doi: 10.1104/pp.18.01552

Figure 4.

Figure 4.

Effects of DkERF24 and DkWRKY1 separately and in combination on transcription from the DkPDC2 promoter and analysis of their protein–protein interactions. A, Effect of the combination of DkERF24 and DkWRKY1 on the DkPDC2 promoter. The LUC/REN ratio of the empty vector (pGreen II 0029 62-SK [SK]) plus promoter was used as calibrator (set as 1). Error bars indicate SEs from five biological replicates. Different letters above the columns represent significant differences (the combination effects were compared to two individual effects; P < 0.05). B, Interaction between DkERF24 and DkWRKY1 in Y2H assays. Liquid cultures of double transformants were plated at OD600 = 0.01 dilutions on synthetic dropout selective media: (1) SD medium lacking Trp and Leu (DDO); (2) SD medium lacking Trp, Leu, His, and Ade (QDO); and (3) SD medium lacking Trp, Leu, His, and Ade and supplemented with 5 mm 3-amino-1,2,4-triazole (QDO+3AT). Protein–protein interactions were determined on QDO and QDO+3AT. pOst1-NubI, positive control; pPR3-N, negative control. C, In vivo interaction between DkERF24 and DkWRKY1 was determined using BiFC. N- and C-terminal fragments of YFP (indicated in the figure as YFPN and YFPC) were fused to the C and N termini of DkERF24 and DkWRKY1, respectively. Combinations of YFPN and YFPC with the corresponding DkERF24 and DkWRKY1 constructs were used as negative controls. Fluorescence of YFP represents protein–protein interaction. Bars = 50 μm.