Figure 5

Electrophoretic mobility shift assay (EMSA) showing the association of MaWRKY26 with promoters of JA biosynthetic genes such as MaLOX2, MaAOS3 and MaOPR3.
(A) SDS–PAGE gel stained with Coomassie blue demonstrating affinity purification of the recombinant MaWRKY26 protein used for the EMSA. (B) MaWRKY26 binds directly to the promoters of MaLOX2, MaAOS3 and MaOPR3 containing W-box (TTGAC) element. Biotin-labeled DNA probe from the promoters or mutant probe was incubated with GST-MaWRKY26-N protein and the DNA-protein complexes were separated on 6% native polyacrylamide gels. + or ++ indicate increasing amounts unlabeled probes for competition.