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

Figure 1.

Figure 1.

Action of DkERFs and DkWRKYs on the promoter of DkPDC2. A, Physical interactions between DkERFs, DkWRKYs, and the DkPDC2 promoter, using Y1H analysis. Auto-activation of promoters was tested on SD medium lacking Ura in the presence of AbA (Aureobasidin A; SD/−Ura+AbA). Interaction was determined on SD medium lacking Leu in the presence of AbA (SD/−Leu+AbA). B to F, EMSA of DkERF23, DkERF24, DkERF25, DkWRKY6, and DkWRKY7 binding to the DkPDC2 promoter. Purified TF proteins and biotin-labeled DNA probe were mixed and analyzed on 6% native polyacrylamide gels. The presence (+) or absence (−) of specific probes is indicated. The concentration of the cold probe is shown; the biotinylated probe concentration was 1 nM. G, Regulatory effects of DkERFs and DkWRKYs on the DkPDC2 promoter. The LUC/REN ratio of the empty vector plus promoter was used as calibrator (set as 1). Error bars indicate SEs from five biological replicates (the regulatory effects of various TFs on the DkPDC2 promoter were all compared to the value of the control [empty vector pGreen II 0029 62-SK]; *** P < 0.001).